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.1./a o q, o g q, T: o -g F ct ll 7(. 4r'"./ce -*.! Qnne.t a)14 i ASL" ft-ts P& l|.! ir ||ld@ k Erdo..d by. ocl,L |{.nLir ItE rudd sdopo grdo Lp €r lrc nEnbr€nd, tu holu, d!€.rlsrlna, dldarddpo.erscd||M adlBolslel Mld Ih€ porB rs t€ eslEiiayslnEr4h *ltdr ool4aE lilro tE ctloddsh dnt nls n.h6 and gorEtc ndeid (mF|{A erb uts ru6! anotF q'rotEm, ibnsrBhd€0€fud€.'dldoog,|l hMB *,rh dmmn' drd r'ot omd !r dm@a ro s .n i b drd'sd fprc 10.6 a Hulrl x rrnoTYtE. Th. ildieidul di|t,]|{'l@ dnr d]f l+ tlE 2, f.in diE rid.ly i'r 14 .!d i! 6!r|rE F ddh I' dljr pr.p.ndd! .h. drM llG bcn {'GdEqlt dii.d @ nd-E dft@ ir dEn ohlEild ud b dlri.3ljri tid dady 6m @ .r.dE, Ndi6 dii ndh.tr of ..tFm D& m Ery rinilrtd n . i.Lditl. 4t t? balX i Ctao,1*. *< /ti,'a3t { Qttr,lT - Are aa-rr ,?r/o."4-ra-; /aa.v- Qoo/ fgure 4.17 MTTOCrtONDnlA. Thc inne. n€mbrone of a nimchondrion is shapedinto folds 6lled itrcr$c th€ su.hcc ate! for oxid.tivc mehboli'm. crosssectlonltrocul micaogliph. tr n I E E tr Opte ltye,<(t I T.eot<.,;Li A t.sl- ll,to eh"lol Fi3!E'6' C.6IrhlonBr!.dbrdrLlDNi{ft.rrx !!BrEIndrsedbyrh.m.k!6 fat*c,it . tu.ddtynlotlhFc!tx^4lddadtalh.lolid.rd!n*.nndn rtd6hlhr n rod..dnd DnA.E hdd.n by ol.r.ordlll b rt. k r.r rli urtsr ldr rh. rmd.u lor rn.r ]n a, q, (D C' >.-> L T's EE o Fg -s o (, .l< o .t-o F€ = olr ,/a/ .!/ru Tabh16.r Fhcnotyp€3.53ocletcd*hh romc mitochondtlal nut dorE Nucl.otlde Mltociondrl.l chrigld conponad.ffi th.idrp.l 3460 NDlofconpler lb LHON 11778 ND4ofGmpler I LHON 11184 NDB ofcomglcr I LHON ESS NARP 3243 MEU5,pEo 1211 MEI.AS 32gl MEIAS 3251 PEO 32fi PIO 58S2 Pto 't03 !814 fiitAct' Enclph.loprdrr E344 MIRNF 8356 MERRI gSC' O'dlo'|lFFthy r0006 NNAC' PIO t2245 PEO t4?09 rRNAotr Myoprth, 15023 htrl infrnlihnMllttitcn dliordlr r5990 Myop.tht tLHoN ttb./s her.dit ry optic n.uroprth, NARPN.uot..ic murcj. s.l,cts, .t tr., atlahls PE|mtot$ MERRF ratodonlc .pil.?.t ..d n$.d.Ed frb.r 3tndrom.; MELA Mitodo.di.l &roPathy. .iaPhaloP.thy, l.dic .cido5i5, itrokclih .P|rod6; PEOPrcgtestive *t€m.l oPhihtldoPl.gh lConola | 6 NAOH d.hrdrcr.na3., Comolq v i5 AtP synth*c. .lnrRilahrru4.ih. n r;nd;for€ith€rA;rc: in tRNAsi'lrcn.th.Y rtandrfo'.inq T o' c. ,lK A tVtra<,?aorp Rt)2. l€Fecn //16€4/ z€/t ? Figur€r5.2 Inhcrltanceol myoclonlcepllep5y wirh raSg€d-rcdRber ditcas€ {MERRI} In humant.{A) Elecrcnmicrograph olan abnormalMERRFmltochordrioncontainhS Daracrvsbllineinclusions. 18 )Th€ pedigree ;hows inheritanceof MIRRF in one famllv and rhepercenkSe of thc r lochondrlalneach Dcrso!found lo bewildtvpe or mulani iMicro6aDhcounesy ol D. C.wallace,Irom J. i"1.sho-ffnlr, M. T. L;t, A. M. s. l,ezza,P. sclbcl, S.w. B6ulngerand D. C.Wall6ce. 1990. Crr 6l: 93r.1 u 15 10 100 100 100 94 94 93 97 96 96 E5 90 Rflf, Cr,v 3e er€o 7a zOtat ftr/ auO../.O Ails' 4f *a, tBl Mitochondrial DNA patt€m lor lndlviduals lndicatedln bluc o o ,1,,{ DNA 4.6 kb II I I 4,5kb I 4,1tb III I I I I All thc prog€nyof a pantcular f€m6lc Y 1.3kb hav. the !ah. patt€m of mitochon&ial Stn€llor DNA bandsas in the moiher, DNA 0.4kb Figurer5.r MalernaI inherilanceofhuman mitochondrialDNA. (A) PauemoIDNA lragments obtaincdwhen mirochondnalDNAis digesredwith ih€ restndlon enzym€pdell. Thc DNA typ€ at rhe leh includcs. fragmenlof3.6kb (r€d).TheDNA typ€ at the righr contalnsacleavage rit€ lor flaell within rhc 8.6-kb fragment,which resuksin srnall€rfragm€nrs of4.5 kb and 4.1 kb (blue). (A) Ped'8recshowinS marernal inheriranc€ ofrhe DNApanem with lhe 8.6-kbfragment(red symbok).Thc mirochondrialDNA rype* lransmhredonly ftrough rhe morhcr. [A[t€rD. C. Wallace. l9a9. Trehdtin Ceaetict5t 9.1 .12af / LodLL.l .ur.eEqtd."r rrid- Pdt-t -d@belfttuflork@imes rtn ntbrd4q TUESDAy,IUNA n, 2&0 oNa Do[r ticCodcofHuman,Ii bIs CrachedbySeientist ASIIARID SlJfrI ? Rivsls'Anmunccfi! iiarts t{srYiledicr tua,Risb md All t nomrt rarra eyl|s$hM!dl.l.l|u! ..i.'dnfuhr.d!, ar. d br drdS ftrr!Fdi-&Ill 0r rLr rr rd..d h r tuq-rdE-Du ?u/t,i h^a * e f'zer O/rrz.<€a.T 9e.47 ea.r&t - (o5t' ft. titlE /<*l*r (aa1'et14 hq/b./ lab 2llerr r 7.t< leJ\--g Vlhat was the the Human GenomeProject's budget? U.S. Euman GenomeProject Funding ($Millions) FY NIHI U.S.Total 1988 10.7 t7.2 27.9 1989 18.5 28.2 4.7 ---- 1990 27.2 59.5 P=l t99l 47.4 s14 134.8 194 2 59.4 104.8 164.2 1993 63.0 t06.1 169.1 1994 OJ.J 127.4 190.3 1995 68.7 153.8 222.5 199 6 73.9 169.3 243.2 199,7 77.9 188.9 266'8 1998 85.5 2r8.3 303.8 Thesenumbers do not include consbuctionfunds, which arp a very small part of z<,r,uoz a?2 2<'re/J? *c1 fe?4*w<,,*< OwZ| /, 3//4cc &aceoan aF 7?€ 6 6.ota24. 4.r'tcaac /*ZfQ_ Frequency(%) Description l,t Tr.nshtedpotions of the 2t,000 scncs dttered aboutth. .hrnmnmm.s t4 NoncodingDNA thd constiNte,$e gred mriority of erchhunrn s€nc 5 Regionsofthe senonethat hrve becn dupliorcd 2 S€qnen€eth$ h$ chmcrcristis ofa scnebut isnot, frnctjonil sene ?0 Constltutivehctcrochromrtin, localized ned cendoheresand telonrcr€s J SrureringrcpsB ofr f€w nucleotidssnch s CGG, tepeatedthousnds ofnn6 4' 2I%: LonginrcEpeEedelemenB(LINrE,'hichrerctive@nsPosons I I %: Shoft intesp*sd el€nenc (SINES),which rc 'ctiv€ @NPosons 8%, RetrotrinspoioN,which coit in longt.nnind repeiE(LTR' d crch end t%: DNA tmnsposonfo$ih 7E ,laaet adt wE t?At O.A fe? 4ocd, ?*eatl aqrce It Oc4 .t lct ca)-.1 /tt zr ,t ' E LINES E SINES Ef LTRS 9.1 A huixn Lryotyp. corulrB of t6 chronosom.l A karlotyp€for a m.h b $*n hel€;a k ryotyp. tor a femalewould havetwo X chloinosoffi //sMlPnobt.t€./ Table20.6 Ave6gecharaderistics of genes in the humangenome ,?ttqae 4.1 leE (aata 347 8.8 ntr/j, Slzeof intemal.xon 145bp Sizeof intron 3,365bp 5lz. of 5' u r.nslat€dr€gion 300bp Sizeof 3' unirdnslatedrcgion 77Obp Sizeof codlnSltgion l,l4Obp 'rotal lengthof Sene 27,000bp q, g a, E o sq, (D s C' E J I ql -s + s oql sU qt q, s(, q= q, o o .t- sq, o ql z g .v,o a .J o- i: o oll -t ,. 4e HArVlv 6:Et/a.tE la.vzityt ,ftrwt,e - -Noocodig t6gm€ a osrwofi genG Conlsadv€t rogarbd ! a€qllonc€o / ngurr 7,t Oc.uEtnc.o{ dlfu hndr of un$l. .nd npeat€dDNA r€gn nt on duqno.omdDNA 7Avteq ?t2C.2,, 22g 4ttFtz &a zpA t/../r<) F2n2ca2-i zig // zt 2.aw at Cllct-+trt7g / Celladheion l5z,l.9%) Mislldeus {1318,4 3%) cyost€leral slilclud por€h (876,2.3%) Tiai6r/ernd Pot€in (203, (2&r,0.9%) T6scdpnon iacror (1850,6.0%) srrui,turitpotiin a mu*telzso, t ov") Potoorcog4. (902, 2.9%) carciun bindhg pot€in 1.0%) Nlcteic.cid enztda (2308,7.5%) s€lect {34, InlEelluld lE rsporiEr(350, 1.1 %) signding md6ul6 (376,1 .2%) S6l*t dgdatory md@d€ (943.3.2vo) $/rnh6e dd syrlh€i.s€ (313,1 .0%) oxido@ductrs6 (656,2,1 %) Moldula. tuncrbnunknown fl 280s,4r '?%) Fi[un l2r0: A tunclionrl cl..tifictiion ol huo Kflownorptsdictodlunctionsfor263s3hu'n.npo|ypsptid€.sncodinggsnss'c|6ssi'icstionis..cc!d'ngtoth€G0mo|.c!|glfundion ;"il;;J;;;;i;il",tgriirctstsene o;to6sictsssmcaft;--sssl^lciroi-836r-0rto-19f:::1YT*::!":ifi":T:"" ffiffi:ffiil;f;iffiffi&;,;;i;;;;iirooirs"r-cd81,r$4-r35r,u'ithpomissionftomthsAmsdc6nAssociationro' thoAdvrnc6msm ofSci6ncs, 6a .4.24 go+r.. t?tAet 4t )alru,/.- /, - ,t/e/tJ o o, a, T' q, o \) ql 3-o q, s oa, o r|F o o Eg o IL {-g ql 3- o q, l< t{- o I T: d olt 41,h,/., lct ?a,vreaa.ec Cibobp?+t g.notyp6 Ab g.notyp. CC Figure2:2 K€y conceptsand tcrms usedln mod€mgenedcs. Note rhar s stndeBcne c6n hav. anynumberol6[ales in th€DoDularion as .awhoL, but no morErhan two alites crn b. prclant ln ary onc indlvtdu.l. &a/riZa-<, .V2 a"*-; 21p )$+c./ a aeltt lt v+< * au aJUr4 I .jo 1- ? %.4 rg..e.2 -; . {- o = T' T's H 3- Jo o ) qto (D {- lto 3- ct () + o a, .o s T: o q, (D olr 'ff 2,Ve l/4an ziar, it 7ztca"/-W& o< Two cystlcfibrcel€ g€nes iren lwo hoalthyindlvidu6h 'f,ahaa\ {ra7" g,o.t.nl !-Zt*.pufi Arows indic€tobaso,pairdifterences Agsrc 9.2 B.rep.lr drfeEn.cs b.trt.6 oNA To be otr th€ safc side, supposeyou assumethat only 80% (0.8) of the 3 bilioo bale par$ /a'i m_|DegeDome ffe noDcodhg,and on averageonly I base A <'a.E palr In./w rspotymorphjc. Wirh rheseassumpdons, you can Qt&. determinerbe ftequencyof polymorphjsrnwirhin a single in_ ar dividual by nulriplying 3 billion by 0.8 and tlten multipling that amountby lr00j 7if,.o^" (3 x to1 x = 0.8 2.4x tor,(2.4 x l0r) x + = 3.4miflion. Theresuk ot3.4 millionis aston ishing: lt meansrbat Uere arE millions of difrerencesberweeD any rwo haptoidsels of humatr ctuomosomes.Combined with differencesin coding and reg_ ulatory sequences(which occur much less fr€quendy), th€ millions ofpolymorphisansat atronymousloci contnbute0o an etrolmouspool ofpotential DNA d|a lels. //p e4au4. /?-/l &Y t?ttl..,t /2 ?/DacEpcct Acy'zz2n 4 Lt; - O, /i4 / 4+..nr. o. sc''l g J d L J gl U s (, o (, CL sO z .9s-q, a .E€.9.8 s b oEI s U|Ut()i o -e^9 .F ts ts o o I FF&< J o € C'.CI UT' 2a .Tat/' {ai. d y'1. lrr.ztaz Cetlantc oa"ri y le7/,J"<>L,, ) gLrung ..q$nc. Orlgh.l DXA l. Iraffldonr Pudnolor Pu n€, Pydmldlnelbr pyrlmldlno (.) B.ro I^lbl ld bl lilhll ld !^llrlEl ld hl rubrtlnrdon hlhl onecod;I-chansed 2. ransv€rslon:Pudn€ bI pydmldlne,Pyimldln€ bt Pudn6 (b) EesC t--.-i}---t (b)O6l6don lnranlon -- \Aninr€dronor. I I delerionatteu the I (c) r,i:i##il."*x*ll hs! ReililriJrlrrfr - d.l.tion \ lT,'.1|'l b*lry.rnn rur.$o.,,|lt b.t srdtlrb,t., rffiorl|.lrdrt Llldl 4*f f,-?, ,R< t7a) 4 .?24'ar@ clauset /A^l ?arc _4 a//Q/ y'lc av? /lq.Ap +_ €eae, oF*o"t, 0'//.rr-rtt t/ *< J7,Yo f,u? o lzzeh A-,q f,yacnce Cl4y/rc T^.BLE 9.1 Fve (|rtsr of D A Polymorphbm Rrte of Mutrdon r{|mb.r Fr Singlebase Muta9ensor to-3-lo-e l/700 bp 3 million /D**) Slippaq€during Io-' r/30,000bp 100,000 9al Minisatellite to r Fewerthan 100 families known,yi€lding t00O copiesin all Delelionr Duplications Tmnrposableelementr (elcludingthore rerulting lrom micrc' or minisat€llite Compler.haplotype Not applic.bl€ Notapplicabl€ (any lo f<..2 a /24.1 Cor" /<./ z1 - /zti./z< Clate.t V?taa 55-2@ - CC€ l€p€at8 OrsodngDM Pt€nuEr.d .lLl.: l}|€l€al€ nol sardl tddorsro cals€ lro€ o-x ryndrcrn€. but lhsytrEy dpErd in t|€ rBl g€r€raton, ffrb |*zm-z,oooceo repds*l 14, Ibh![]Xdrfilaon|.h-dadwtd|.d|.rd..td. w cor|ELd.t| (htf..lb) 6d. tsl3:tr! It&/.b ur{rrtodJ l?.10 rh.COO R.prr'lntl.flrF, G.n E9.ndwlth E dl lea-/ .ti Table18,1 Examplesof geneticdiseases caused by expandingtrinucleotlde repeats Numb.r of Copl€tof R.pc.t Dls..r. R!p.!t€d s.quenc. Nonnd RrnSe Dl3€.5! R.r!. Spiftl andbulbar mlstular atrophy cAc 4o-EZ 50-1500 lecobsensyndrome ccG r00-1000 SplnocerebelLratarie {s.v.rat typ.3) cAc 4-44 Autosomaldomlnant cenbeller stixl. 7-19 37-2ZO cTc ilzF300o ffi CAG 9-37 FrlcdBldr atad. CAA 200-9m Dentatorubrdl-paludoluysl.neroplry CAG 49-75 t'ffdmut.plLpiy of th! z-3 lz-t3 ,(ttat /a 6 {1) (cGG)5 5 5' 3 3' (2') ,^^^, 5' 3 (cGG)${.o 5' 5' I ,.\/\/V\-r\/ 3 (cGG)>roo 5 3' ,r\/\,f \.f \J\-f \.f \,r E Dltoa.saauslngall€ls n n tr Hsl.ro2)!ou3 or h€mtzygoo8 >200 >200 EguE a Amp|tflcrtlon of rh. r.tpt n r.tci (cc Gond't., *fth th. fr.gn x ry.dro.n . (1) 8egionot rn x chrcmolome containinga oormalFMR-I gene wirh 5 rcpeatsof the CGGseq!€nce on onenr.nd. (2) FM8-I 9en6 in unaffededp€opte g€nelatt hav€ tutrerth.n 50 FFars; unsiabteprcmut Uonalets of the g€nehde beth€$ 50 and 200 copierof the repear.Futtbtown di*aleoustng allekshaw morc tnan 200 CGGEp€atr lome mutlnt a el6 hde moE than 4000. (3) A fBgite X p€dtgrceshowing rh€ numbs of CcC r€p€aBin dir€rent individuats.Note that Indivtduabaffected bv fradite X tndrcFe aE dmon alwaysthe progenyof mou€rs who cr;ed- ,4arn 2oA./ o+4 lteeti ra^t ti .ta./rnr2u u( f 'tFt* tJ.al 0 ,hat;r2oJ.-,,^l 4 Peee /t/tttt Anclstral fia.lllslte .hromolom. I I DAIm RSlp -/ f.lrtC Ztcat pe6on6b dlg€6ted V I E E - I I gl.ph of the g€1. A8 ThbeEmpl€ .sutrr€r th.t Bobis homozygourlot theA patErnand Jo€ i5 homozygoG fo. tle B pattern. A p€En lEcrcrygousfo( tne nn wluld dlspLy ban(|sre6n in boththe A andthe B p.tternr, lt :t arHdo|| n|!|llJ|tlqdr ?.Unr.tibn rrt.|rlk d|-i.n l||.t c.r i. ur.d h Ertttv rTr/o.lo. /.rZ ,(R?r (ev tt at€o 7. Zalrb./aab 2o,/ tal tB) MltochonaldalDl{A pattem lor lndlvldualr indlaatedin rcd b'.fi:' 8.stb 4.5kb 1t.1tb All th! progenyof a Dardcularfemale '1.3kb hav. thc samepsttem of miiodrcndrlal Smrll€r DNA bandsas ln the mother. 9NA 0.4kb Fisurer6.r Malcrnnl inheritanccot hrma n mitochondrial DNA (A) PaiternofDNA fragm€nts obrairrcdwhen milochoDdrialDNAis diSestedwhhlh€ retlriction €nzymetlatll. The DNA lypeat rhclelt includcsalragDrcnl oI E.6kb (red).The DNA rypeat the riShrconlains a cleavaSetitetor ga.ll wirhin rhc 8.6-ib hagmenr,whtch resuhsin smatlerlragmenls of4.5 kb and 4.1 kb (blue) tB) Pcdi8re.showir!8 maremal inheritance oI the DNA Panernwith lhe E 6-kb I.68ment{red symhdlsl-Thr milirhondrial DNA type islransmirkd onlY rhroughthe moih€L lAlkrD C wallac!. 19a9.tehds inGett.tits5t9.1 hrla.lon/-Z *ee*br 71< dt4<-a.J .tt;oo Pcn <4( ,{ ar€t 2 a.*r/V (2lr ie/ a*tf ztci ti (a) 0 ,,,1'l'1 tu. fu1ar* 3' s' o . (b) atl€rdis€suon f35o Allele1 froo will E*rr Lr'o Rourt 9.7 t.trtlGtlon .h. PolymorPhlsnt 1. Oetomines6qusnc€s flanking micmsaiallnes /SQpav /? 4+r 2.Allplit allelesby PCR L".14s ==3 :- --a- ry91L. ,TO 9= 5 $aa-7 7k.Qqt lot bitlucls 3.Analyze PcB poducls (b) All€lespresent in populalion Qfto ra /0e1rry Diploidg€notypos presenl in population 'll1 /a./i ty'u<(r 212 313 112 1t3 23 3 2 1 rlguE 9.r2 n€tetlon of [email protected]€ PothotPhlrm! by P(n .nd 9.1 d Y. Fiiu.€ r7.r2 C,cn.ricvarirtlon in a VI{TRuscd lr-DNAtyplnS.E€ch numbercd lane co ains DNA hom s slnglcpersoD the DNA hasDeen de.vcd wth a r€stdcdoncn4rme' scparac! bv .lectoDborcsis.alrd hybridizedwlth radlo- a&tvcprobc lxl. ttc bncr labclcdM contatn nolccller-wdgh nrrlcn. lcourcry ol R. W. Allcd.l @ //r.4 la L+r'tf /.r/",,;, 9*t- ?as-4 itn;tt SIR,l 15,16 15,16 slR-2 8,8 7,r0 STF.3 3,5 7,7 STF-4 12,13 12,12 STF,5 32,36 11,32 STF-I 15,1615,16 15,16 srB-2 8,10 7,8 a,lo STB-3 5,7 5,7 3,7 siR-4 12,13 12,13 12.13 STF.5 11,3211,36 32,36 2Lrrr/.2 .feueeat u'va(tzn) 1atF.v.Tre) Dig6slDNA to oblsln minissl€tmer.agm6nrs_ rndividual lndividual BC lr* I t; r*1 {-J l--J _t-_\ ffiffifl* tlltll t{,e. - rt<./n -m: -E== ffi S -5TFry ft lr't.Zy' RuntEgments on a get.pgrfom Southen btor, Hybridi2ewi|n pob€ containinq t41? minrsqtgtht€ I ztt/a)tlaJ ABC t] 4oy...l a 6 5 3 tlgurc9.r3 Mht.|tettft€ .n.trtt prevtde5. ndB of .lmuh.ncoul|y d.t€ ,oa- z?a&* &./o-/n ,+ ka a/ ,/Vd?t/2.ta ?/,.i = r/4tc 'Urr* 7lu/ea /<2..4 z5 z"rly' " 8..1 trt'{*2p./y u&-an*l} I . !-L (X|.3doi: How Samolecollected at a scarnofcrlm! Susped Suspect2 EEEq) @# w& ONA DNA DNA I Mlcrosatelllt€I V V sequence\ 8 repeatsof cA r€peats repeatt /2 /8 irt.fffftftmft.jn7I riffil ft- =:Effi ffi I roPcR. ;templat€ V t r.rrrrrrrr.rrr.rr/l:i'l' EF Effi :r r.r. rrr[..r.rrr Prlmer -iEi- ..ffi ffqr*"r, Template DNA I I I I t numberoi .opiesof the ffi ffi mi(roFreliirer€qu€nce. lt@ Thet69nen$ ares€parated .II Ditfe.ent.rize f ragments appearas difiereni bandr. :l collectedat the(ene of rhe €imematchet DNA from ffiffiffi Resultsof oneSTR locus Multip'emkrosarellhe loci on th€gel. Conclu3lon: Thepatt msof b.nd! produ.cdby d,ff.r.{t rrmptdsare @mpar€d. The bloodrt|ln rp€ qt 19.:fo DNA flna.rprfndnt c.n b. dto ld.rntty. p.ts. [6d cou,ety of oftttad c. !n , A4nnantovn. Ma tyl an d. l h*; f,q..nlzet Zr;J /.a // aa.c €3i ;EE -rar = r o-iH ------._._.= = E, =\.-7'll--- t === a r! -; = -- - : *E----:- -=!: - :': a FigureiT.r3An exampleof DNA typin8 in a criminalcase. Each pan€] is lhe res lt of DNA typing lor a diller€nt VNTR.Thelanesmark€d Sr,52, and Sl coniainDNA from blood samDl€soI th r€emalc srspccrs;rhose in columnsUl firough U7 containDNA from semensampl€s colleoed lrom seven femal€vicrimsolrape. Th€ lanesmark€dMconbin molecular-weightmarkers.In each case, rh€ DNA from suspec52 marches!h€ samplesobGined lrom ftevidims. ICourtesyofSlevenJ. ReddinS,Offic oI lhe tlennepin CounlyDistrict Alrorney,Minneapolis, andLowellC. van Berkom and CarlaJ- Finis,Minnesota Bureau of CriminaI Apprehension.l V.t-m /tcf 4rz./ - h,z,v 24e..+,1 4.qa .Qyr.)tf @ +z/|/7ti 2ot1 /Q?tr7?)ry ee /n /rbttalr 4r tfio biologicalcommurities exposed ro chemi- cal, biological, or physical stress.They are also used to monitor geneticdiveKiry in endangeredspecles Eptdemlology and food ral€ty 6clence. and speciesbred in captivity. DNAtwing alsohas imponanr applicarions in trackingthe spreadof viral andbactedal Evolutlonary gcnetlcs. DNA polyrnor- epidemi(diseases, as well asin idenrifying phhmsarc studiedin an effon to describe the source o[ contamination in contaml- the patle$s in which different types of natedfoods. genetic variation occu! throughou! the genome,to inler the evolutionary mecha- Human populatlon hlstory, DNA poly- nismsby which geneticvadation is main- morphismsare widely used in anthropology tained,and lo illuminatethe processesby to reconstruo the evolutionaryorigin, which genetic polymorphisms wirhin globalexpansion. and dlve$ifiaation of the humanpopulation. speciesbecome taistormed i o gen€ti{ diflerencesbetween species. Improvemcnt of domegtlcated pliants and anlmals. Planrand anlmal breeders have tumed to DNA polymorphismsas geneticmarkers in pedigreestudies to iden- E tify, by genetic genes mapping, that areas- * sociatedwith favorabletraits in order to incorporatethese genes into cuffentlyused I varietiesof plamsand breeds of animals. Hirtory of domesticatlon. Plantand ani- E mal breedersalso study geneticpolymor- t phismsto identify the r4ild ancestorsof cultivatedplants and domesticated animals, as well as to infer the practicesoI artificial E selectionlhat led to geneticchanges in A patern thesespecies during ity t€sl-for win€s,cen€tic res€archers domestication. haveshown rhat the nodon ot "puriryof breed" DNA polyrnorphisms as €cologlcal In- rhouShtto be so criticalto rhecommerclal value dicators. DNA polymorphisms olcen6inwines is ill-Iounded. DNA rests are being indlcatelhat winegrap€s such as rh€ cabemer evaluatedas blological indicalors of genetic sauvignonand chardonnay (shown here) arctn diversity in key indicator speaieipresent in facthybrids. 2.7 Appli!,/tiot!' dD'',7| Iiottut @ Foacorra. (taa4.4 bL,/t,k ,zl..J.lc-/t* ) Cra,aa2;, ar.--lU2 C2... ttll ,tl 2/seqr. {ert €t 7 (9 'RFtfs 4t b, tt 4,r& h 4Lt1ct-:L:?aRo/' Locui ot , "dl3ot!6 gono" l! genltic rilk t ctor,, O ONApolymorphlsms (g.nstic mart€ralalong Figur€r.29 Conceptsin geneticlocalizadon of gen€tlcrisk fadors for dlsease,Polymorphic DNA mark€rs(indicat€d by the venlcal linet that areclose lo a Seneticrlsk faclor (D) in the chromosome tcnd to bc hhcrtted rogelherwlth th€ discaseltself. Th€ Smomic location oI th. risk la(lor i5d€t€r- nined by ex.minlnS th€ known Scnomiclo.atlons ot the DNA polymorphlsmsthat arc llnk€d with lt. ilriQ*all' rt* aari*t t t4 {alSBE|g.n. tcl G€notyp. S€od t ONA PhanolYps ooa%, a I Dnpl€x tn 18,SBE|gsno with lmenion lmutanr''f tttle.t.wl D auJ Fieur€?.2(A) lt, (round)ls an allele ofa geneftatspedRes th€ amino acid sequence oI st.rch b;nc[in! enzvmcI lSBEll.{Bl w iwrtnkled)is an alleletharencodes an Inactlvelorm ol lhe enzvmedecauie iE lHi sequencels interruptedby lhe Insenionol a lnnsposableelrment. (c) At rneievel of the mo1fiffip6Al phenofypc, w it domlnantto wl Genotvpewwand wi' hav€-round seeas.whercas cenulluii rvwhat wrinkled tecds.The moleculardlffercnce between lhe allelescan bedcrecred as a-restiiZton fragmenl lenglh polymoiphlsm (RFLP) using thc enzymeErRIand a Drobc ul hvbddlzesat theshe shown. Al ftc molenrlarl€vel, thc allclesar€ codominanl: DNA iron cachgenotype lelds a dilferent molccularphenotypc---€ singl€ band differing In slzefor homozygoustywand ww,and both bandsfor heFrozySousl'rw. Cttc /al aa 22a Tvlryt * 4.r4: ott)otttq/ Qfrie ,(n?t t 4aa*att taiovrtrl Ve lE&le atc. 4aZola *"$d G€nomicDM lrom: SicklecellFglobin gene 'r33i3133m'- lll.* Msttr Mutaltofl sttr witr Msttr Ilomaladuh FSlobin gene 5't*:lFiFFffi3 r L:-:i_--i-:l tir Southemblot ttl Itlsttr MsuI sul - Hybidization pobe Figur€ ZL7 kection of the sickle-cellhemoglobin mutation by South€mblot .ntfFis of $rromic DNA3 cut with r€6Eiction€nzyme Mtttr fce ,. Gt?; A a/..-r1t ,kaaplrl^ ,l 4/1"6t nguE 9.1t Dl.gno.l. ot h.rnophlll. thNgh tft. Indlrlct ikt 142bp- Aclls{6 l- 99bp I aa.ir" i €bp I P.s36nt _T Uti/ Al&k - S.ali a/2at,octca fha/t/2 f7../a zr/1c&t /. ;/ 6c*c (b) 1. Heallhy 2. Heallhy ;ffi:; :; -f-:: :.-i iit'--t-;3:-;: :.-l:: PCBamplificalion or allsamples ngur.9.9 Udng FCr Utth AlO. to .Lttnli. q€.|ot'F .t rltti pgt"Un t*-. (u) sefoE Perfoming lhe genoVPingP|otocol Y llir is neso,y ro synttresUemo otigonucl€tids that differ at oot a Dsnab6,f,'ys e e sinqleba$; oneof theseoligonucleotjds is comPlementaryto the tttl the ri.lle<€ll t,ttrtl wiE-type F{lobin allelq the other is comPlementaryto m allele,ftese two synthetrcDNA molecules sere asthe ASOSfor lhe V VVickl*ell genotypeassay. (b) GenomicDNA emples obtainedlrcm .-.-----.-r/ indMdudlpeopledrc tubjecled to PcRamPlf(alion wlth Pdms \------@npl€mentaryto nonPolymoPhicsequen.es that nankthe ba* that mutatesto causetickle-cell anemia. (c) Theamplified emple ftom dividedinto two aliquotrthat a€ blotted diFcdy to tc) I @ch indtidlal is t ,r,>q..:. filter papei (d) one aliquotrtom 4h emple is hybddizedto the 23 123 1 wild-typ€A5q the oth$ aliquotis hybddizedto the tick!+ Hybddizewlth lab€l€d 3;l['ffN'8i""* I Y I ldl r -! ' t, '-t .'.:,: tit, *l-.-t;":-: l'-',',i't, ;"|';l .ffi['-ili'l-";';>l (el wast' or rnnyorioizedprcbe and exPos€ to Jilm n-o--_-ll-TJ F 4rn1 lrlt fi&qi €co.t I3' 1. Bipo €gqsarc €ln€v€d 2. Enracl€d €ggsa€ 3. At6-10c€llslags, lrom lh€ ovarywith a sydnge. isnjlzsd wilh sp€m. on€ c€ll is Emovsd llom sach ot six viablo 3mbryo6. 4. In 6achol six isolalsdc€lls, sits ot commonmulaton in CFgene is amplfli€dwfih PCR. 5. DMd6PCR Droductlnlo lwo oodlons.D€natu€. Applyon€ dot ot €achsamd€ onlo nlirccollulos€lllt6r. U U I I 6. Hybddiz€ior 7. HybddlzgwthAso (b) Dlrgnode Nomal ASO MuranrAso ' cslb llom €mbryosla!8r nansplanred inlo orarus f,g|rr 9.r D.t.dr|g d-.tdc fbrodt gdotyF of Gmb.yonl. cclk. (a) ,, ttrefeniliation and pEimpl.nt lim dbgnoCa(b) C.ll 2 ig homozygourfor tlE nonml .lldq cel ,r ! h€terc4gousfor the CFmltation. 3- u,I os .F o3 T' 'Ea, ,ri -€> 9ot qr -s o+ !.s oi =o vr6 PE EP ar> rE v, q, o !) -s : \FI z o z () a ct lt q, T' C' = q, T'o = a, o 'rt 3o -so .Fo o= o, AL Gro. <-a, +- lt .8S F9 T: az BREAKTHROUGHOFTHE YEAR HumanGenetic Variation Eqdppedwifi lsstol,cheaporlechnologios loi 8squsncin0 DilAandas56sing variation ir gonomoson scslos rarging fronono to nillions ol basotrosoalchals arcfinding out howtruly difforEfi we arsfrom ono snodtor TTIEUNVEILING OF THE HUMAN GENOME ALMOST 7 YEARsAGO castthe frs fainllight on our complete gen€tic mrk€up. Srncetheq cach (.6lE:D nelvgenome sequenced and each nery individul studiedh'l illuminatcd ourgenornic landscape in evermor€ detail. In 200?,researEhers camc to apprEciat€the extent to whichour genomcs ditrer fiom p€Fonto perlon c-6-Tl andthe implicationi oflhis t€riation for deciphering ihc g€netics ofcom_ ple,(drseas.s and personal trdits. variation Lessthan n yearago, the big newswas lrianSulatinS ot !nlqu..chanqetin gen€tic whlt n!l.t betweenus andour primatecowins lo get a bctterhandle on thenumberand orderolgen€s {A-D) :l changesalong ihe evolutionarytree that led to humans.Noq $e have addvadety lothe homan genome- mov€diiom arking*hat in our DNA makcsus humanto strivingto knor wharin FryDNA makesme mc. v.l 449118octots 20o4a.&bEvffai6t ltrutt ARTICLES A secondgeneration human haplotype mapof over3,1 million SNPs The InlernationalHapMap Consortium' Wo dss.rlb€ tho Pha!€ ll ltapltap, wfiich ch.racl€titos ov€r 3.1 million human3lngl6 nucl€otlde polymonhlsms (sIPi) .ohmn sNPvrrbrbn h Sgtovp€d In 27OIndlvldual! ftun fo{r Soogrrphlc.llydlv.rs€ popul.tldls and inElud€s25-35% of ire popul.tlons luw€yed. Tho map i! €sttn.t€d to captrre ungp€d conmon veri.tlon with tn .v€ra8s m.ximum I ot belws€nO,9 and 0.96 dspsrdlnSon popul.tlon. W€ d€morBtr.t€ that tho ortgtt 8rnorttitn of comh€rcbl SBnons{ids Sdtotyping prodrcts captur€scornmon Ph.se ll SNPswith an .Ysra8emaxirnum I ot up to o.a h Aftlctn rnd up to O.95in non-Atri.an populatlons,and thrt pot€nti.l grins in pow€rin aslo.lrtlon studl€scan be obt.ln€d lhrougft imPut.tion.Thss€ d.ta aLo rev.rt nov.l .Ep€ctsot th6 structllrs ot llnkagedlsequillbrlom. Ws showtftat 10-an9t ol ptlrs ot Indlvldual.wlthln a popul.tlon shareat losst dr€ t€8ioo of utsd€d 8!n€tk kl€ntlty .ri3ing ftom .s<6t alc6try and th.t uPto l% ot aU conmon varianB ar€ uitaSlEble prin.dly bocrusothet ll€ wlthin rsGomblnatlonhotlPots. ws *or that teombln tion rlt€s vrry 3ystornat'rcally.rdlnd 8t!l.3 .nd brt en Sglet of dffddlt tunctior. Finalt wa dqroGH. inctd€d dfffsrlnuatlon at noo€yndiynoul .ohpar€d to synonymous,SNk, r6ultina frofi 3Ftsmatl. dfhrglcr3 In th. strcnSlhol .tfic.ct of h.turrl .6Lctlon hwen populltlons ,f4./P.r Et aq2 EQgauVZ It'sAllAbout Me Along$ilh lhe tlood oI discoveries jnhuman g€neti6,2007 lawthe binh otanewindunry: p€lsomlg€nomjct. Dep€ndingonyourbudget, youcan eitherbuy a rorghscan ol your genome orhav€ th€ whote thing lequenced. The companies 5aylh€ inlormation vfll h€lpcunome6 leam aboutthemg€lvesandimprove th€h heellh,8ut egearcher5 worrythat theseseMc€t ooen |lo a Pandora! box of€thicatirtues- At5300,000 toslmilLion per qenome, *quencing all3 bitlion base pairsi5 nilt too (only for all but a lew.Allhough dozens mote personal genomerwitl prob.bly be sequenced in the coming year, mon llbe doneby pubuc.nd Fivrte research organjzations-jnctuding lheinsti_ rut€run by qenom€ maverirk,. (raig Venter, {hote peronalg€nornewas oneollhrce.omptetedinz00Tintheljniled5t tesand china. In a torer budgetetlon, Hafl ardl 6eorge Church this motnh sitl deliver jnitial DNA requenc€ifor the prol€in-coding tection! (10/o of the genom€) totht fid 10volunteers forhi5 PeBonalGenome Ptoied.llletnvlhile, i negl com' panycalledl(nom€ i5oflerinq full-genome sequencing to 20 ('lnometr filtlngto pay 9350,000. Agtimpseolone'sqenom€halreadyvtithinlh€reachofordinarypeo_ pl€,lhank5to severalcompani€s. Th€yinclud€ 23andl$e, $hich has financjnglrom Gooqle and may lel $etslink to olh€rewith tharcd traitli Naviq€nics,whid vrillscreen for about 20 medjcal conditions; and deCoDtGenetic, in keland,a pioneerin dieeategene hunting. For 11000to12500, lhese comDani€, wjllhavp (onsumetr send in a saliva "SNPchip{tos(anlheit sampleorcheekswab,thenu5e DNAtora5many as1 millionm.rkeB.lhe compani€s willthen match th€ retulte silh the titestpubtcations ont6itr, common diseaset, and ancenry. Allhoughmany cuslomerr may viewthig €xercire a9a waylo tearn funfacts aboul then5etver-rerrealionat qenomic, rome calt it- bioethkirtr.rerary. Most commondi5ease na*er iden_ iril;j tiliedto lar rairerieks onty tr11 5lighlly,but they corld cause needlesrworry. Al thesame time,eome peopl€ may be ter ified to tearnlh€y have | rcla- tiv€lyhigh dsk for an incunbte diseaseguch as Alzheimer'r. lhe rushtovrard perronal genomesequenct a$0 snap- enrlong-h€ld flor esabolt prelent discrimination.AbiLlto P.dorr'r bot?rhls deek-ffrab kil insurergand enptoyerr lrom (ouldrcveal youi inlimale r€crcts. misusinqgeoet'( data is naled inConqrcn. Complkaling maners, your gen€trcinionnation exposes yout Themon prolound inpLicatione of having onel genome analy?ed naynor be whal it reveabnow---iehich isnt mucHnt whatit maythow lareron. Perhapslo rid€nep such questions, some companies willli'rit whkhmarkerr to dirdore. otherr. houever, willhand cunom€Blheir €ntiregenetjc jdentily, atonq with all thes€crets it may hold. 'locllYll lAlsli ocA2 Fmm SNP€di! OCA!' rh. @ulocuLr6u dbinim B.d. (.le loowdaln.bulmPD6r.irB.n,or,DNl0),ir a g.d. ei.r.d wirb albirirm rd etuin pi3D.nblio! .ff.cts ir gd.nl $cb 6 .y. color, rr&dr ocn2 0n|*/d*rdpdj.i4ldHocr) -' !-r* d*"a;r^,r A 14. (>3,0o individu.b) dudt of Can6id Gd.xlt m1[$//*w.dlJndji]L3ornB/!e? indiaB ttlr lt. tollo*i4 OCA, veiuts, dl lqlkn i! th. fi's itrtu of lhc scrc, e pr.ls. ially linkcdto bluc.yc colorinn nu,@; lt9-/ww.rlij'in.r'ib.!d/sNsip-nf .qi? rog.th.., tb.y fom b.plorypBl[d (i. sm. cec (hryJ/lw.ldi,ol[,nh€d/,i6/q@? .r l.do pftdid cy. colo.*irh 8fter thu 50:50 PrbM.d 4943 odd!.IPMID l?236130;OMIM m32m.ml3 (htDr/ww,rcbi,nln.Dib.gov/cntE/dkp.mim,cgi?iG203200&.=2(B20O-AU.I.Vriotm13)I Th. hlplotyp.r e d.lio.d ii ord6 a lislcdabov. lor ln.* 3 SNP',$, lor .rmpl., $. TCT h.tbryp. rcl.6 to u7451?4G)-r64tr,263(C)-Fl1355019(I).Thc cmrF coccb.tv..o diplotyps (lh.t*o hiploryp.! h on. individu.I)dd $. % ol i ividudr witn bludgiy, gE thev ard bEwD.y. @lor , spdiv.ly, *a Ep.n d a follov3 for thcnort 6mmor diploDTclPMID l1Z361301: . TCT/TGn 525.2&0.9.5 I Tcf/lrc:47.l.mi.32.6 . TGT/CCI 24.6.l4-3. t.I .'IG TGC:n,9,22,1,9,0 . T<{DTC| 25.0,43,6.7 . tlfnti.20.1.31.0.43 r TCT/mt: l?.6.38.5.aa.o ' TGT/CIC: ?9.233.68.8 Thr haplotyp.sshoen in ,otd t rri6 rcptMl tncons Epon d by lh. d$oF of tnie3rudt io b. rEd |!Bi&n wi|n bdn ay. color,FunnmE, $. h.plotyps ,bosn abov. @ d publih.d, .d 16. .'&ia|.n SNP' - ehich h.v. ,ine cbsg.d , 6 s.ll - @ ootin tb. od.nt lioo shovoh dbSNP, MoE *c.ody, a dudy of . ldg. Ddi.h fdily l.d ro .si idr ei|} 2 SNPci.. difi.EDl Egjo! of OCtt a liol.d b blnc6t bswd .y. olor Edi.r n i.s fonodditLcot E8i.trr of tb. OCA2 3.d. ro rl$ b. pr.nidiv. of ay. color; . oCAz SNPBlsrxxol icrp. prdict bFwo cy. aloi IPMID 12163334PMID 1588p0a6;OMIM [email protected] 6np:/tusw.Dcbi.nln.nih,sov/.okr'dispomim.csi? id=2032!0&e2620ojlLlicvridtool l) I I OC.l2 SNPulgx)4{t nay b. |!ei .d wi(h tEr^el .y. colori. somcp.pul.liool butnol dtrn. IPMID l2l6333rt PMID l$49045; oMIM 282@dll2 (f,trp:/tu{w.mbi.rlDdh-iov/cDr./die.mim,cgi?id=2c1200&.4r,2m_A .licvriEor2) l humaneye color. 2007:23andMe introducesthe first Personal Genome Service. Unlocktlresecrets of yourown DNA. Today. welcome to z3andMe,a wcb-basedservice that helpsyou r€adand understandyoul DNA. After providing a saliva sampleusing an at-hom9kit, you can useour inte.activetools to shednew light on youl distant ancestors,your closefamily and most of all, youself. Glllwn r'" o"wat aistrdMe Gene.Iournal JaI,22,2@8123andMe now availabloin Canadaand Furope. Jan 18, 2008:23andMelaunches its blog, lhs spittoon. Whatdo your genessay aboutyou? rx( GeneJoumal G6mJoumal GettingStarted \trith 23andMe Order Form First Nsme Lost NaDe You haverot addedaoy kirs yet. Click the b OrdgrSummary Kits in Order ?'utt*,t l"t* 0 kits 7rlr.7 Pdceper Kit httpst/Uw2l.ndn..com/ou6.Me/pG.33/ $999.00USD 2randM. - OurS.rylc€:H@ th. Prd.rsworks U 2Al0A2:29 PM |TP <1f-r 4at fctv tu2s Spit Kit f36f 74< GOu,ve Joining23andMe is easy- onceyou've placed your orderand signed our onlineconsent form, all youdo is spitin a plastictube included with thekit we ship to you. Eachkit is labeledwith the nameof the personit is designatedfor alongwith a claimcode. Just use the kit's pre-paid, pre- addressedshipping envelope to sendyour sampleto our contract€d laboratory. GenotJ,Tirg After receivingyour sample,lab professionalsextract DNA from cells in your saliva.Your DNA is thenchopped up into shorterstrands and copied maly times via a processcalled amplification.Next, your DNA is washed overa smallmicrochip-like device that contains short strands of synthetic DNA. The syntheticDNA fragmentslatch onto the piecesof your DNA that area complementarymatch, Then a laser-scanningstep reveals which strandsof syntheticDNA are stuckto your DNA to determineyour gelotype. Thechip usedin our processis thelllumina HumanHap550+ BeadChip, whichreads more than 550,000 SNPs (single nucleotide oolymomhisms) plus a 23andMecustom-designed set that analyzesmore than 30,000additional SNPs.What this meansis that the laboratoryprocess rcads nearly 600,000 datapoints on your genome.Find out moreabout our gglElypilgIleegss. hnpt://lw.23.ndme. rqdwr diiaEdsNPh'reur.dat ::W, Birhip3 en L.h i, iit ,dtins pEie i,ms orene. c QCI/n-*e /k.,taa. a7 €t112.r1o.7r$ ,LraryJ €avt 6.vzgt_c - -.-- GenomesforALL Ne xt- ge n e ratio n te chnolo gie s thatmake readingDN A t'ast,cheap and aidely accessible arecominginless than a decade. Theirootential to reaolutionizeresearch andbring about the era of trulypersonalized medicine I meansLhe time to startpreparing is nou) Fra2 o.'4 hait-t-:2. ,)""7 -.4a2 4e, v 4q7.; { a,*, .tuJ. z2adt/ ou,aGSTrT:g €ac.yoc.rratC lmport nt dlvsloDsrt In Oll,ll $quonclnr Mi6rch6r: Dilcov6r6d DNA Avery: D€monstrat€dDNA a6 "Gsnsric i.latsrial" Efficloncy lvatson & Crlck DiscovsrodDoublo Hsllx Slructur€ bp/per.o year Holl6y: S6qu6nc6dY6ast ni|AM . SpecilicRNA diSestion and chrcmalogGphy methods were usedto sequenceRNA;it reqrired IaGe quanlities oi sample. lvu: S€ousncodr coftsciv€ 6nd Dl|A . Pimedsyn$esis concept and 2.D electrphoresis were used; sampleswere labeled and less materialwas rcquired. San8or:Dsvslop€d didoorylarmlnadon s€qusnclngprocodun; GilborrD€voloped ch6mlcal dslradadon 3oqr6nclng protocol . Chanleminabon a_d chemicaldeg.adation roncepts '^,ercdeveloped. . Polyacrylamidegel e ecvophoresiswas used to separateoNA lracts. U$3lng: D6volop6dMl3 cloniflgvsctors . Cloninesyrtem was appli€d. Hood:Oorclopod parli.lly afomaiod s6qu6nclng3yit6m . Sequencingreactions were optrniled,. Assorted sequencing strateges wefeappliedand comput€r assisteddala handlin8 rvas slaired. Vsntor: First baclsrial t$om6i r6qu6nc6d . Automatedlluorescent seque(cinB iistruments and rcbolic operations rvereappledtolhe process.. PCR sequencing concepiwas introduced, Effcl€ncy bD/rnachlna/y6ar Pgrtins-ElmorCorD.: D€vololsd and mark€tsd 96-crpillarylaquoncsr . FulyaLtomated96{apillary el€cirophoresls sequencing system is availableto researc h laboratodes. Figur€ 2. Advancesin DNA sequencingefficiency and bor-iniensive process- Howevea finly automated sequenc- someof the te(hnologicaldeveloprnents that enhancedthe ing machines have now largely replaced human s€- produchvity of sequencers.Initiatly, all the stepsin DNA quence6, greatly inc@asing efficiency. s€quencingwere perfomed manually,making it a very la- tao4l _ /xtaP/2/,L.- eR /x aa/"72..-. - k ttacl ni . 4,yre Crrae.r/ay -a d -aI r- a /ttno.// d*a a4z?ta-ftt .l4e 1V22aat*6/ 140,000,000,0@ 120,000,000,000 100,000, c E :10,000,0o0,000 20,(D0,000, 19851986 19871988 1989 1990 1991 1992 19S3 1994 19b5 r Sge t sbz r ggsr 9b9zcioo zcior zcloz zobg zob+ zo'os zdr FICUREIO.I Crowthof DNA s€quercesd€posired inro rhc pubtic daraba6es(CenBantq EBr, DBIL) fiom lgOS to 2006. uenEankand rheorher pubtic darabases beSan their operations_stoing and diss€minating most ofthe DNA sequenc€savailabl€--in rheearrv r98os. The increas€ in sequencedata resurts from the ;ssjve increasein sequencint capabilitiesthat cam€ with th€ HumanCenome proiecL pafticularty in tfie tate1990s when liql:!:ilg ot the human genome began in eahest. A mitestonewas ,ea"h"d in zoos ,rh"n m" 100,000,000,000th(one hundred billiondr) base pajrwas deposited into th€ databases. f.gure 18.4 ATJToMATEDsEeuENcrNG. Thi! ,cqucnccFcility srmunncoury ruDsEuttiple .uto@rcd !.qu.dc.n, ..ch proc.ssiag% s npl€srr r ti@. karu /i- 2,v,4 t€?o;4 @ az q, s +- r{- o q, g -9 \) o # sqt E c'r Ei: tLI te O) DM hbeinSand stind dbs4i'iih {g R€adiryte *qu€n.. fromdE aub..doratl Molecrleto t€ *quft€d (many(opies) offi9n9n9ffi 6 c c+T u*' L r*. ,,*'*"n*l* I Dimebvlsulhl€ I r DrilsoeoPc I TT I offig-q-qffi I !l I I =t affinriffi = I =I offign9nFffi ME = offi9*qm9* I ASaoie8el Pipeddine I qffi9 -- -) Purilyon€ ol GffigfrFfr! FiSure48 chemicrldeSradation sequencing. 4f fPfttre r'arar t?'lulut14, /n' / /a f2 zt rzi 21 2,v,1 f,, n /tl.*i J?/aroe i 'll.tlECrl|rflAlrd C!5t!|rLtEabtttd 'IrF DMeald 6tt8 rlshtt*l4b th€BnCab b.t€ svnfE* catE co(tlokrsrtdy Filttd ta| h golllrg d fErdt&l*1. h @.Ut l /rthq- l/ tY..a ft^z a1L @ o..2 1.vz2s I CH Dideoxyribonucleoside triphosphate(dNTP) triphosphate(ddNTP) 19.25 rhe dideo,(y*quenclng rcaction reqolE3 . rPt i.t sub3tEt tor Dt{A synthe3ls. {a) stru.turc of deoxyibonucleoside triphosphatethe nomalsubstEte fof DNAtynthelis. (b)Structurc oi didsxldbonucleosidetnphotPhate. wiich l.cks an oH Sloupon the remplate-3,IinmFfiFaiF[m - Primer-5'HH y itrmr'n I r,rrorrm \ + DNA ' 6,"a"d on"ti,p" "f I dideoxyribonudeordeI viphosphare(ddNTp) L -+dd^Tt I ffiryffiffi\ r ifl' P \1,,t,/\ol{,< t' 5' ! q 1at a./ the iragmentsprcduced li each leadionare sepa€ted by gel - I e n..-.j i i Auroradioeramor G C electrcphoresisqel Ihe sequenceGn bercaddnectly cc tom thebands th€t appea. on theautorad ograph of thegel, staningirom the bonom. 5' 3' Thesequence obtained s the 'Sequefice compement oi theorginal Sequenceof of complementaryorigtnattemptate strand st.and 19.26 rh. dld..r(y m..thod ot DttA sequen.ingh baled on ihe teminrdon of DNAsvhthesir. C4nylti , -S ,/arba.li / DNApolymslass -- DNAtolym€|Bso - .i TCG 5'-a TCG T G TCGG TA C TC TC TCG lof ddC j; TCGG TA C TCGC TCCGT TCGGTA TCGGTA 5'-A TCCGTAc T 5'- 5'- A TCGGT A TCGGTA CG TCGGT TCGGTAC GT 3 T G c G c T G c T c T G G T3' c Sho er T e Igure 1 /.1 t ]\{{NUALaND AUToMATEDENzyMATrc DNAsEeuENcrNG. The sequenceto be determinedis shou at the top asa templatestrand fo. DNA pollme.ase{ith " priner arrached.a In rh€manual method, iour reactionswere done, one tor eachnucleoude. For example,the A 6rbe would contain d,{IP, dcTq dCTP, dTTq and ddAIP This lerds ro iragmentsthat end in A due to the dideoxy terminatoi The fragmenrsgenerated in eachrezchon are shown along wirh l}le r€suitsof gel electropboresis.,. ln automatedseqtrencing, each ddNTP is labeledwith I differentcolor fluorescenrdye, whicb allows the rcactionto be donein a silgle tube.The fragmentsgenerated by the reaclionsare shown. Wle. rheseare elecrophoresed in a capilla!_vrube, a l.ser at dreboftom otthe tubeexcites the dyes,dd eachwill emire diff€rentcolor that is detectedby a photoderecror tn't 14. (t,le ,l2ti - ( It.l, E ltthD{ hi C!r,rllom [.thlo E tfi Th6CM dard d |lE l|0ft &ll4 b IEffi€ hIE 8yrfiEbOf TE dddilditry stad tr€t b gro^rtg dm bi bato. t{ '* "*--t{ ?2t' /e/cea/ a,lut Zp* aalJ. f1r2lt /rer/g. /.-J ,f€Qaa"zi PF "a,feaaG -fte Figur€4.9 Pyros€quencing. strand sndresis rcaclionis c.nied olt in he fifrTillTrmTn,,,,, ,,,, absenceof dideoxynudeotides.Each deoxynudeotideis addedind'vidually, along wih a nudeoiidaseenzyme that degrades + d|e deoxlnucleotideif it is not incorporated dAn + degrads into tle strandbeing synhBized. Incopolationis detectedby a fr.sh of + d'emiluminesc€nceindLrced by the dm -) deSrades pyrophosphatereleased from lhe deoxynudeotide.Ihe order in which + deorynucleofidgale dddedto the growing st'dnd cantherefore be follow€d. dGIP ldemiluminese.re= mniiiii'llrf,,,,,,, + dCP + de8Bd€s + dAn l.hemilumneenre= ffiinnrfinfi,,, ,,,,,, + o- Ql/ea /.t >9 I R-t AHPLIFICATION 8...u..liehtia8n.l..r.ditficuhrod.r.ct.rrh€ecrl.of.ein8l.DflAmol..ul., b.....xr.nsion orliiarionr.aciiohs.r. ott.n p.rrom.d onmillion6otcopis orrh. s.m.t.mpr.!.svand sinuhaneousls.C.ll'n.. 6.rhods[c ridb] ror m.kinqrh.s.copiosinvolv. PCion a mini.ruriz.dscrl.. t:\ rl Polongcluior OPoloniespolgm.r.secoloni.s-cr..t.ddi..crlgohrherurt c.of.3lid. or8.l€.ch.mt.ihap n.nshich.lchpl.refi aem.ntcanfindrdbindio.PcR sithih...h polongprod!..s. clu€t...onrainingmillions of r.nplar..opi.s. Temolat. ODropl.t'.6hr.ini^tpolgmerferathih . 6n6il.molsionc..s.rv.aerinqPCR -l .h.mb.rsro produ.e b..d pob;i.s. when ...mplarerr.smmtanach.dro I ab'adkadd.droechdropl.r,PCR orr.,rlurt- p,oduc6rum on(.pr.sor0. r.mpl.t.,.ll.na.h.dro ih.hcad. IIIJLTIPLEXIIIG Sequencingrhousandsor6illions oti.mpl.t. tragm!.t6ih parallclmaxinizes speed.lsinsl.'soleccleb.s...!r.^sio^ sgtr.musingtluor.sc.n!sign.l d.t.crio^,lor€rampl.,pl.ce. hu.dredsotmillions otditt.r.ntt.mphi. tr.gm..tsonaringlc.ii.slb.,owl._fil.Anoih.rh.thodifrhobilir.smillions olb.adpoloni.s dn a r.l sorf.c. forsimuh.n€oG3.qu.n.in8bq ligaiion *irh nuor.sc.nc.sign.l.,shown in rhein.g..!.iqhr bolow,which r.pr.s.n!! 0.01perc.nt otrhe iot.lslid. ar.a. lr! i, a..i ii )ttri.i Czarvm7 f,lra rn-/.a ,/ t;... /xtao/p/,(- ,(- ksta /ilae faatM hrr2 ,,tf tt]eftes FaR t&/i< A442/nart ilil il t'f il| I t ' At rlrl, G€nohewidearrociation nudies are In addinqto kn6wnetdches ol DIAconn€cred tI:litilill'|,il'lt withlype 2 dirbd€ekolored bars). SNPedia . New modelfor prostatecancer based on 5 SNPs . rs1815739sprinters vs enduranceathletes . rs4420638aJltd rs429358 can raisethe risk of Alzheimefs diseasebv morethan 10x i rs6l52can prevent baldness . rs9939609tdggers obesity . rs662799prevents weight gainfrom high fat diets . rs7495174green eye color . ts1903146in 3Voot the populationgreatly increases the risk of type-2 diabetes . rs12255372li\ked to type-2 diabetesand breastcancer . rs2395029asymptomatic HIV viral load set point . rs32,1650influences intelligence and alcohol dependence . rs1799971makes alcohol cravingsstronger . rs17822931determines earwax 7k l+co-./ d, DNApioneer Watson gets own genomemap fitF nilgenonE of JatrEs\tibbon x'io jd{y d!covq€.|tle lhrcin! oi DNAIn 1053.he3 !€€n d€ctlfE|rd, mor8ng$tut |drt. ad€nllab bdld/s b t|e gate 'aYb 3n hDr|dlng €ra ol pe$dlalEed gBnorflc A co9yo{ hi! g€nofiF, t€cold€do.r lvio ovD., r/l/6!or€s€ri€d !o \.',ebonon rhrlday h 3 oo€nlonyIn Ho|don DY or !|e Humsr Rlchad Glbb!. dlr€cto. Jmr{b\o{ffic,xoM|'.6('dd G€nomeS€qusidng c€nt6r d 0|. B6ylo. C.lhgs of .dldr€, and bYJomnBn Rohb€rg,ftt'r'6r ofth€ cofip€ny45a tlb 6 PtlnEr_Friendly oG E 3 C.lumn F.mat TE6late ll shaeArdde 6t Lisbn to Artde 'l arn hdled lo !€6 fiy gEf|dr€,' \lv.lL.l 'T r.xtga - + Rohb€Efs.oop.ny lnal(€3.n tuDvilit'e ou ..i "n"tti .iam, t" tots.t v€.slonor d {h provd caPabl€ordecodlns wbLon'' p|rtbnl a.ficrE h tr tr|.rrh E brt tEn at n hn a€ldMlctEd Eghoh' i'lt to' ;rrrn .t4fr. Irb !.q'rt. sE! |gfr€d *! andvzedbv Giltd' cdb h tlddon n w.r GiDb rfio rftlo..d [r lr.s otlql€tdng ! illod! gcrd![' aen Qna,zc -fc aeala /-z !,!!rnome" Knowthyself. Knome is the first personal genomicscompany to offer whole-genomesequencing and comprehensive analysissewices for individuals. Based in Cambridge, Massachusetts, we work alongside leading geneticists, clinicians and bioinformaticians ftom Harvard and Mtr to enable our clietrts to obtain, understand, and sharetheir genomicinformation in a manner that is bolh anonlrnous and secure. We are cunently offering zo individuals the opportunity to paticipate in our initial launch phase. By being amongst tlrc first itrdividuals in history to havetheir whoregenome sequenced, these participants will help pioneer tie emergingfield of personalgenomics. Recerrt Neurs January22, 2oo8:Knome Commences Whole-Genome Seouencing process For Filst Clients January 10, 2oo8: Knomeand the Beiiine GenomicsInstitute Enter into Exclusivestrategic Aliance November 29, 2oo7i Service for Individuals /zto, ,,oo U/.1 a"/t .llu 4 l/, A 4. hwF2a /v- 71.; T' oJ = H o {+;l g o IL a, E o sql c'l € q, U E'o $E sfi o g$i: (, rts H ct lt 2" r?rc. ..itil,?ifpRrzE Looin / Reoistar $10Million to nE Firlt T€omio s.q6tEs 100Human Genomes 10 Days a,,az !e/ aSS,/aa2,4 /rnl,1 Lrth.( "The 1000Genomes Project will examinethe humar GeDe-sequencingprojects keep gerome at a level of detail getting bigger. tllat no one has done Tuesday,January 22, 2008 before,' said Richard By Emily Singer Durbin,Ph.D., of the Wellcome Trust Sanger Institute, who is co-chair of the consortium."Such a In a testamentto the steady project would have been plummgtin sequencingcosts, unthinkableonly two Ye3Is todaythe NationalHuman ago. Today, thanksto GenomeResearch Institute amazingstrides in (NHGRI) announceda massive sequencingt€chnology, intemational collaborationto bioinformatics and sequencethe genomesof 1,000 population genomics,it is peopleftom arcuod the world. now within our grasp.So we are moving forwad to build a tool that will geatly expandand further accelerateefforts to find more of thc geneticfactors involved in humaohcalth and disease." uunng ns two-yqu production phase,the ltOU GenomcsProject will deliver sequencedata at an averagerate of about 8.2 billion bascsper day, the equivalentof morc than two humangenom€s every 24 hours.The volume of data--andthe interpretationof thosedata--will pose a major challengefor leading expertsin the fields of bioinformatics and statistical genetics. The 1,000volunteers will be select€dftom thosewho participatedin the HapMap project, a map of commongenetic variation(see "A New Map for Health"),and will include: Yorubain lbadan,Nigeria; Japanese in Tokyo; Chinesein Beijing; Utah residentswith ancestryfrom northem and westem Europe; Luhya in Webuye,Kenya; Maasai in Kinyawa, Kenya; Toscani in Italy; Gujarati Indians in Houston;Chinese in metropolitanDenver; peopleof Mexican ancestryin Los Angeles; and peopleof African ancestryin the southwestem Udted States. 'e,, lpE (een Qae,ucrt 2 'l.largs ONAclon8s arc llrsl lsolatod,Th€86 sro conllguouss€qu€nc€! bas€don ova.lapplng lagg€dsn6s, tragmBnt€dInlo smallor clon6l lor s6qusnclng, 3. Th€ snlirc s€qu€ncols ov€dappinglargs. don63. 1. Cul DNAol 6ntrc cfircmosomslnlo 6mall irrgm6nls and don6. ray -.+ -==-::_=---=-- 3€gmsnland anEnge bas6don ov€dapplng nucl€olldos€quencas, figure18.5 coMPAxrsoN oF sEetrENcD\c MtTEoDs. a. Th€ clone- by-cloft Dethod u.er ldge dones assenbl€diDro overlappiDg regiols by Sfis. OD@ ssmbled, th€s€ @ b€ fiagmetrt€d iDto sDall€r clonesfor scquercir8. ,. In th€ shoqun method the ertiE g€none is Frgm€nted into sndl doaes and scquoccd. Conrrter.l8oridDs $s.mble de 6n l DNA s.quoe b.scd otr d€rhppitrg lud.otidc s€quenc6. 74a tst&r r Q-r.-e 1r 4t22.. :---- - DltAlraSmdte | &ib!e 8€l + ele(|rotld€sis tdb'r-rr.d' UliEl: s..iohd t i,,fr,s,rce0M tr.rdr4'.rr.d' UIt 2:0M ndrlcrt ;'i ir l2 It.d|dh.!db 'i*-1 .l purityDMhom ! Utesd - - - Y'"1't'IlR'D- 'ar**r-dE, ft€parca donelilEry a| '..ia::':...;,..7 I obbin€nd5lquenes t ol Dt.lJlinsat .| Figure4.t o lfie wayin whichthe shotgunmethod was used to obtainthe DNA sequenceof the Hdenqph,i,usinfluetuae Eenome. H. influerzoeDM wassonicaled, and fraSmenbwith sizes ben/veen 1.6 kb and2.0 kbwerc purified iiom an a8aroseget and liSatedinto a plasmidv€ctor io produ€ea donelibrdry. End{€quences were obained Imm clon€staken fmm this library and a computerwas used to identifoo!€daps betw€en sequences.This r€sdH in 140sequence contigs, whici wercass€mbted into the completegenome sequeflcq as shown in Figure4.t L fue 6qnv.r $orn.r1 / d.t/e Estihrtcd G.non. Sizc (Mb) Eltih.t.d NuDh.r orG.n.r E @ ruF d, 4uit""'"*".:*':"T::::;:.Hl*:"":,:.:,:::"*:11,,.",, ffi @ ffi ffisi6r ffi ffiW W @ qze\ rll nn(n nr rtr ,lr!r nr ln,tr oi lfirr Ccfont EXTREZOrmlrr Frri*- - .**r'lE " no nr errq ";"al I Clear Genome ectsstatistics ) Orgrnism Complete Drrft assembly In progress total Prokarvotes 621 g. 416 1563 I 4 30 83 Bacteda 578 456 1tao Eukaflotes 22 I34 t'72 3A Animals ! 8l lzl0 Mammals z 2! 21 46 Bids L 2 1 Fishes l 9 b!E!E I t9 E. I 3 4 Roundworms I 4 p t7 Amphibians 2 z Reptiles I I Otheranimals 1 t8 25 'l Bb!r.: 2 3.t q I-and planls 2 5 24 3l 9199!41res 2 ! 9 Funei !s 52 D. 91 Ascomvcet€s c 20 7l Basidiomvcetes J. z 4 12 Olherfudei I 2 5 8 Protists 6 l8 27 5l AdleEdereE -! 9 L tl Kinetoplasls I 2 5 8 Other prolisls 4 7 25 total: 649 594 64t 1891 Revis€d:Jan 28, 2008 bl.t 4u b. ?a a,r,nl /lc( aF 7/:r.- ltze 57A?e.,.6t,2 figure24.8 LIVINC GRiAT APES. ' Gorllh | '#S'iss: All living Crearapes, with rhe excepuonof htrmans,have i haploid cnromosomenumoer 3f not losta ctuomosome; 5I fisu.6lz28: Sarcoding ol p.imrtochrohosoE.s 63 a wayol nvo.lirg nruduld dilloroncos Cros-speciescotor banding {Bx-Flsd) prcliles show aliqnm0nrof o.tholosous primato chromosomes with human chromosom€s l_22 and X.Chrc'nosome s€G (wilh numbe.inq accordinq tothe human homolosd showllom lolt to righthuman, ch mpanzos, qorilla, orang-utan andmacaquo.Toimprovecompaisons.chro'nosom€s2p/2q,i21,l4ll5and20/22,whi.haiesinqlechmmosomesinhumano.inthe nacaquoaro shown rogelher wirh ths g€at a9ohomologs. Thistype of anaLysis has sugqest€d a prulimmary ancesl6lk3ryotvpe lor humanand srestares Eeproducsd rrom Mull€r a nd Wienbers (20021 Hirm 6erel109,8F 94 with pomislion itum Spdnservs aq lAot I ka rl . U.1., 2t-, .1r.2, A Qaatq 4 ryeute ? o - I I =i!=,'=i- I == r=-t -= l- ii+!!E:i:12 3 4 5 6 7 8 91011 !li12 13 14 '15 'iE16 17 18 19X HumanI I lrll rlttlltt tll r T 12345678910 111213 1415 101718 192021 22X (b) Sp€cl€s1 o O F|GUIE12-t0 Synt€ric relaliondips betwe€n8€nes in differentspecies, Mammalian genome, arc chaEcteriz€dby ta€e chromosomalsegmenb in whicha f€w up to s€veralthousand genes, spanning hundreds of lhousandsto millionsof basepairs, are in the sameords andoienhtion in distinctspeci€s. (a) Each of the 20 mouse chromosomes(aulosomes l-19 and X) h coloredaccording to the chromosome(s)in the humansenome withwhich it sharessyntenic blocks. The key to thecolo6 of th€human €hronosomer (t-22 andthe X) is shownat the6ottom. For example, th€re is a larSeblock at th€rip of mousechromosome 1 that is ortholo- gousto a s€gmentfrom human chromosome 8 (y€l/ow). These regions were part of thesame chromosome in the lastcommon ancestor ot humansand mice. The complete synt€ny between the x chrcmosomesof miceand human beings is striking. Th is is charactestic of the X chromosomesof placental mammals-all are derivedfrcm the sameanc€nlal chromosomeand therc is grearevolutionary pressurc to ke€pX-€hro- mosomeg€n€sto8erherand not mixed in withautosomalg€nes. (b)Syntenycan be used to id€nrifyrhe lik+ ly odholoSot a protein.The arrouE represent 8enes. Wh€n the sequence of 6/',egene in Species1 iscom- paredto thesequence of theentire g€nome of Species2, two genesin Species2 are found to matchit arthe fevelof 80% sequencesimilati\y lgny cicle andhexagon). A similarcomparison is donefor the neighbor- inggenes. One ofthe b/uegenes in Species2 isfound to beflanked by th€same neiShbo ng genes (DroM and ye ow) as is the blue gene in Sp€cies1. In conttas! the secondb/ue gene in Sp€cies2 (hexagor)is flankedby two genes(orarge and rcd thathav€ no similarityto theflankinS genes in Speciest. Theloca- tion ot the b/uegene between two similar genesin Species2 makesit a verystrong candidate for the odroloS of the b/ue genein Sp€ciesl. Caap'tra /u) -16;aeac,(1 e-, a., et ,/ E. coli FIGUREl3-I7 Thecircll genomemaprof 0157:H7 E colinaine K-12and Ol57:H7. The circle comparedwith depictsthe dGtibution of seqlencestpecific K-l2 to eachstrain. The coline4 backbonecommon to bothstains is shown in blue.Thepositions orOl57:Hhpecifi. r€qu€ncesare shown in rcd.The positions of Kl2ipeci0c sequences areshown in gren. Thepo5itions ofOt57:H7- and K-l 2 specificsequences at the same locationare shown in tan.Hyperuariable sequencesare shown in puryle.lAftsN r perna "G€nome era , Sequenceol Efrerohaemorhagic Eihetichqcali A151:H7: Natuft 4A9 20A1 7529T533. Counesy ofcuy P lnken t t and figure24.14 Atrlcln d.€plng llckrll!! CO; TPARATIVE GENOMICS Chagls disease,Aftican sleepin8 sicldes, and claim millions of lives in developing nations each yed, share6200 core genes. Drug developm€nttargeted at proteiro encodedby the sharedcore senes could leld 6200sharcd @rc g€nes a siryle EeaFnent for all t I Taq€l lor drugdovolopm€nt fl. ft/ao 4rvt2 l.r.y<'/ +r ,Q" / tltPLtFtcaTt0N 8.c.!s.li8hi€iF.l....di{liculrrod.t..r.trhG.c.l.ofrsinll.0ilAmol.cul!, b*..dt...aon o,llir{on r...iio...r.oird P.ttom.don nilliod oaclDld oi!h..rm.r.df l.t..tr.nd.imuhrn.ou.ls. C.ll,tn.m.thodt lo.nd 6l lor m.rdn3rh... cotl..invdtv.Pcno. r nlni.i!.i2.d.c.1.. \ rl tolo.gclurt.r O Polonl..-polsr.r...colonid-cr..r.d dn.ctlgon ih. tuLc. oI..lidc cr.l.!cncont.in.p n.nwhich.tmpl.r.lr.F.nrc.nfrnd.ndbindto.PtR uithin..ch polongproducd. cl!.t.rconl.inlng6allion.!tt.nPl.t. $p4... atr.rPCi r.npht I r-i"fr"-:f !oropl.r.coni.lninip!l!m.i8.{hhln' .^oil.mul.lonc.n..d...rin!PCn _ ch.mb.r.roproduc.b.rdpolonid.Wh.n I .r.mDl.r.ft.8n..r.!irch.dto I .b..di..dd.dro.shilroFl.r,PcR orr.ni,o,on t.mDl.t.,.ll.rr.ch.dro lh. b.rd. 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JQae,vcnZ Ze (ea*z.r €xVaZt %nnnb- tt /o1l fa$.|/e,/ NATIONATGEOGRAPHIC NEWS mFomNcYouR tnoflD DAIIY I' NEI/I,SHOME ANTMALNEWS ANC'EI{T VIORL.D €NVIRONMENTNEWS CULIURES llammoths to Return?DNA Advances Spur ResurrectionDebate ior N€tjonal Geographic Ne\i6 June 25, 2007 Todaythe only placeto see woollymammoths and peopleside_by-6ide is on 7ho F/,hlstoresor in lhe movies ad r€.€lrchdsa€ onth6 v€€e d Piedngtogeh€. condeb g€ttom€'or brE_ oea rpeoersua o neanoo.tls andmn|lmtE {s€ea bdefovervlew ot So no{ ihe Ug $adon b, wll r€ sooi D. aue lo bdng3u.r! odnd lp€cl€3 bad( to frb? R€seaftruB .€ dM&d 0l€r ho{dl€ niglt ty lo do tli! sr{ lNh€lEf lts evenbslDle {Rdat€d:1/\'oollY Mamholt' R€.un€Guon 'Jlnssh Palt named I,AId 8, 2dlq.) . ,eS. Z 4. 6. /1, Riday,J.nuary 25, 2008 Synthesizinga Genomefrom Scratch Scientistsssy th€ results rcgesent a newstage in syntheticHology. 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Elch e0droill|E B a l^3llfio' lndh,ldual€oornbinarn clo{|€6 16.11Cor|ltlc-thg r OonaUbrary Hutnanctvofilo6omd DM b bolat€dard hol(€n up Intohagrnfite uslng€sfblion €ruyrn€5.The n4rn€nt6 ar€lAsstod ilto !€cds (plasrnkb6rs shottl hq€) ard hk€n '.F by hod betahl o€b, dl ol |rfiidl fl4l h€rbo€a dngl6 faOrsn ol fb hrtEt onfA TfE ffi.rnatitt in fE tsdtiE bet€tbl cr,itJrs ard !# ol colonie const'hrt63a gd|e tbray. U4. <.tu4 ,&te y'r