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United States Patent 19 11 Patent Number: 5,333,675 Mullis Et Al

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United States Patent 19 11 Patent Number: 5,333,675 Mullis Et Al US005333675A United States Patent 19 11 Patent Number: 5,333,675 Mullis et al. 45 Date of Patent: k Aug. 2, 1994 (54) APPARATUS AND METHOD FOR 4,312,835 1/1982 Zoltan et al. PERFORMING AUTOMATED 4,335,620 6/1982 Adams. AMPLIFICATION OF NUCLECACD 4,358,535 11/1982 Falkow et al. .......................... 435/5 SEQUENCES AND ASSAYS USENG 4,362,699 12/1982 Verlander et al. ... 422/131 HEATING AND COOLNG STEPS 4,395,486 7/1983 Wilson et al. ........................... 435/6 4,404,845 9/1983 Schrenker ........ 73/61.1 I75 Inventors: Kary B. Mullis, La Jolla; Larry 4,474,015 10/1984 Christmas et al. ........................ 62/3 Johnson, San Jose; Richard A. Leath, 4,478,094 i0/1984 Salomaa et al. ....................... 422/35 Berkley; Timothy J. Wennberg, 4,483,823 11/1984 Umetsu et al. ........................ 422/63 Mariposa, all of Calif.; Louis M. Mezei, Madison, Wis.; Joseph T. (List continued on next page.) Widunas, Freemont, Calif. FOREIGN PATENT DOCUMENTS 73 Assignee: Hoffmann-La Roche Inc., Nutley, N.J. 171140 5/1984 European Pat. Off. 0171140 12/1986 European Pat. Off. Notice: The portion of the term of this patent 0325763 2/1989 European Pat. Off. subsequent to Aug. 13, 2008 has been 2650593 5/1977 Fed. Rep. of Germany ........ 357/87 disclaimed. 2413708 12/1977 France . 2161815 1/1986 United Kingdom . 21 Appl. No.: 21,624 OTHER PUBLICATIONS 22 Filed: Feb. 22, 1993 Techne brochure for Temperature Preogrammer Related U.S. Application Data TP-16, Dec., 1984. 63 Continuation of Ser. No. 709,374, Jun. 3, 1991, aban doned, which is a continuation of Ser. No. 899,061, (List continued on next page.) Aug. 22, 1986, abandoned, and a continuation-in-part of Ser. No. 449,136, Dec. 11, 1989, abandoned, said Primary Examiner-William E. Wayner Ser. No. 899,061, is a continuation-in-part of Ser. No. Attorney, Agent, or Firm-Davis Hoxie Faithfull & 833,368, Feb. 25, 1986, abandoned, said Ser. No. Hapgood 449,136, is a continuation of Ser, No, 833,368, Feb. 25, 1986. 57 ABSTRACT There is disclosed herein a machine for performing 51 Int. C. ........ G01N35/02; G05B 17/00 nucleic acid amplification under computer control. The 52) U.S. C. ...................................... 165/12; 422/116; machine utilizes any one of a number of heating and 436/50, 935/87 cooling systems under control of a host computer 58 Field of Search ................. 236/46R; 165/12, 27; which directs the heating and cooling systems to heat 364/557; 422/116; 436/505.5; 73/116; 935/87 and coola reaction-chamber-containing heat exchanger (56) References Cited at appropriate times in the process. The reaction cham bers are pre-loaded with the nucleic acid(s) to be ampli U.S. PATENT DOCUMENTS fied, a thermostable enzyme to catalyze polymerization, 3,311,303 3/1967 Moyes ................................... 236/46 3,392,914 7/1968 Nienstaedt ... 236/46 specific oligonucleotide primers, and four different nu 3,856,471 12/1974 Winitz ....... 23/253 R cleotide triphosphates. Also disclosed is the process for 3,912,913 10/1975 Bunting............................. 236/46R the amplification chain reaction implemented by the 3,983,363 9/1976 Alter ......... ... 219/521 machine, which utilizes a thermostable enzyme. 4,008,048 2/1977 Hellemons ... 422/138 4,206,872 6/1980 Levine .............................. 236/46R 57 Claims, 19 Drawing Sheets 5,333,675 Page 2 U.S. PATENT DOCUMENTS Studies on Polynucleotides-Total Synthesis of the 4,504,733 3/1985 Walsh .................................. 219/521 Structural Gene for an Alanic Transfer Ribonucleic 4,517,160 5/1985 Galle et al. ........................... 422/63 Acid From Yeast, By Khorana et al., J. Mol. Biol. 4,518,700 5/1985 Stephens . (1972) 72, pp. 209-217. 4,534,941 8/1985 Stephens et al. Automation of Microliter Plate Chromogenic Substrate 4,544,436 11/1985 Clopste et al. ...................... 219/385 LAL Endotoxin Assay Method by Use of a Modified 4,554,839 11/1985 Hewett et al. ... 422/63 Pro/Pette Express System, Martin et al., J. Parent Sci. 4,555,486 11/1985 Bahl et al. ... 435/91 Tech. vol. 40, No. 2, pp. 61-66, Mar.-Apr., 1986. 4,582,788 4/1986 Erlich ..................................... 435/6 4,582,789 4/1986 Sheldon, III et al. .................. 435/6 Saiki et al., “Enzymatic Amplification of B-Globin 4,598,049 7/1986 Zelinka ............. ... 436/89 Genomic Sequences and Restriction Site Analysis for 4,679,615 7/1987 Levine .................................. 65/58 Diagnosis of Sickle Cell Anemia', Science, vol. 320, pp. 4,683,194 7/1987 Saiki et al. 1350-1354, Dec. 1985. 4,683,195 7/1987 Mullis et al. P. A. Martin, et al., J. Parent. Sci Tech, 40:61-66 (1986). 4,683,202 7/1987 Mullis . Forma Scientific Advertisement, Analytic Chemistry, 4,685,081 8/1987 Richman . Aug. 8, 1982. 4,708,886 11/1987 Nelson .................................. 422/72 Cole-Parmer Instrument Co., 1985-1986 Catalog (com 4,711,851 12/1987 McNamara ... ... 422/63 piled 1984). 4,737,462 4/1988 Mark et al. .......................... 435/253 4,800,159 1/1989 Mullis et al. .. ... 435/172.3 Lake Shore Cryotronics, Inc., Review of Scientific 4,853,332 8/1989 Mark et al. ... ... 435/252.33 Instruments, Jul. 1980. 4,858,155 8/1989 Okawa et al. ... 422/63 Zymark, Advances in Lab Auto Robotics 1984 pp. 4,865,986 9/1989 Coy et al. .............................. 422/63 1-17. 4,889,818 12/1989 Gelfand et al. ... 435/194 Panet et al., Studies on Polynucleotides J. Bio Chem. 4,933,146 6/1990 Meyer et al. .......................... 422/63 vol. 249, No. 16, Aug. 1974. 4,965,188 10/1990 Mullis et al. ............................ 435/6 Histomat advertisement, R. Jung GmbH, Oct., 1980. 4,981,801 1/1991 Suzuki et al. ..... ... 435/290 Techne Brochure for Temperature programmer TP-16, 5,008,182 4/1991 Sninsky et al. ......................... 435/5 Dec., 1984. 5,038,852 - 8/1991 Johnson et al. ....................... 165/12 P. S. Martin, et al., J. Parent, Sci. Tech. p. 63. 5,075,216 12/1991 Innis et al. .............................. 435/6 Techine Ad for Dri Block PHC-1. Biores, b.v. Bioexcellence TAG-Polymerase and Am OTHER PUBLICATIONS plicione Kit Ad. Techne PHC-2 Ad. Barber-Coleman Co., 1980. Techne PCH-2 Temperature Cycler Ad. Tachine Brochure, 1982. Techne Flow Coolers FC-200 and FC200 and Dip Techne Brochure, 1988. Cooler RU-200. Tecam Dry Heat Baths, Techne Catalog 7051081, 1986. Techne Tempunit or Tempette Immersion Circulators Techne (OG-1 Black Digestor), Catalog 7051091, 1986. Ad. Techne (Ori-Block 08-3), Catalog 7051101, 1986. “Studies on Polynucleotides-The Linkage of Deox Dialog Search for Techne Patents. yribopolynucleotide Templates to Cellulose and Its Use Brookfield Test Chamber. in Their Replication” by Panet and Khorara, The Jour IEEE Transactions on Biomedical Engineering, vol. nal of Biological Chemistry, vol. 249, No. 16, Issue of BME-29, No. 8, Aug. 1982, pp. 557-568V. J. Anselmo, Aug. 25, pp. 5213-5221 (1974). et al. "Programmable Temperature Control System for “Studies on Polynucleotides-Hybridization of Poly Biological Materials”. deoxynucleotides with Tyrosine Transfer RNA Se "Amino Acid Analysis System' Rev. Sci. Instrum. quences to the r-Strand of d80 psu DNA”, by Miller et 51(7), Jul. 1980. al., J. Mol. Biol. (1972) 72, pp. 503-522. "Studies on Polynucleotides-Repair Replication of "PC Application Ideas' Instruments and Control Sys Short Synthetid DNA's as Catalyzed by DNA Poly tems (Oct. 1980). merase' by Kleppe, et al., J. Mol. Biol. (1971) 56, pp. Techne TP-16 Operating Manual May 1984. 341-461. Techne TP-16 Operating Manual Dec. 1984. 5,333,675 Page 3 OTHER PUBLICATIONS linear double-stranded DNA and supercoiled plasmid Smith, M., Appl. of Synthetic Oligodeoxyribonucleo DNA, Gene Anal. Techn., 2, pp. 89-94, (1985). tides . pub. in Nucleic Acids Synthesis. Application to takura et al., Synthesis and use of synthetic oligonucle Molecule Biology and Genetic Engineering . Nucleic otides, Ann. Rev. Biochem, 53, pp. 323-356, (1984). Acids Research-Nucleic Acids Symposium Series No. 7, Yansura et al., Studies on gene control regions IX. The pp. 387-396 (1980). effect of hypoxanthine-substituted lac operators on the Koster, H., Introduction, pub. in Nucleic Acids Synthe lac operator-lac repressor interaction, J. Mol. Biol, 133, sis: Applications to Molecular Biology and Genetic Engi pp. 117-135 (1979). neering... Nucleic Acids Research-Nucleic Acids Sym Goeddelet al., Studies on gene control regions VI. The posium Series No. 7, pp. 1-4 (1980). 5-methyl of thymine, a lac repressor recognition site, Agarwal et al., Nature 277, 27 (1970). Nucleic Acids Research, 4:9, pp. 3039-3054 (1977). Panet al., Studies on Polynucleotides . J. Biological Gergen et al., Filter replicas and permanent collections Chemistry, 242(16), 1974, pp. 5213-5221. of recombinant DNA plasmids, Nucleic Acids Research, Khorana, Research Proposal Submitted to the Division 7:8, pp.2115-2136 (1979). of Biological Sciences, National Science Foundation re: Rossi et al., An alternate method for synthesis of dou NSF Grant, Prop. No. GB-368812X, Grant Prop. No. ble-stranded DNA segments, J. Biol. Chem, 257:16, pp. PCM/3-06/57, sub. Oct. 11, 1972, funded for the period 9226-9229 (1982). Feb. 1, 1973 through Jan. 31, 1978. Szostak et al., Hybridization with synthetic oligonucle Maniatis et al., Molecular Cloning. A Laboratory Man otides, Methods in Enzymology, 68, pp. 419–428 (1979). ual, Cold Spring Harbor Laboratory, 1982. Zoller et al., Laboratory methods: oligonu Wallace et al., The use of synthetic oligonucleotides as cleotide-directed mutagenesis: a simple method using hybridization probes . Nucleic Acids Res., 9:4, pp. two oligonucleotide primers and a single stranded 879-894 (1981).
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