Conventional NZYTech Restriction Enzymes: Activity Guide Enzyme Activity in NZYTech buffers (%) Buffer Buffer for Restriction site Methylation Reaction Enzyme provided for double Isoschizomers (5’ →→→3’) A (green) B (blue) C (red) U (white) sensitive Conditions 100% activity digestion AscI GG ↓↓↓CGCGCC NZYBuffer C 80 100 100 100 NZYBuffer U * PalAI, SgsI (MB231) BamHI G↓↓↓GATCC NZYBuffer B 40 100 (<20) 100 NZYBuffer U * No None (MB064) BglII A↓↓↓GATCT NZYBuffer A 100 (100) (60) 100 NZYBuffer U * No None (MB065) CciNI (NotI) GC ↓↓↓GGCCGC NZYBuffer A 100 80 60 100 NZYBuffer U * NotI (MB153) DdeI C↓↓↓TNAG NZYBuffer U 25 25 25 100 NZYBuffer U No BstDEI, HpyF3I (MB236) MalI, BfuCI ♦♦♦, Bsp143I ♦♦♦, BstENII ♦♦♦, DpnI G(mA)↓↓↓TC NZYBuffer C 70 50 100 100 NZYBuffer U * (MB078) BstKTI ♦♦♦, BstMBI ♦♦♦, DpnII ♦♦♦, Kzo9I ♦♦♦, NdeII ♦♦♦ BfuCI, Bsp143I, BssMI, BstKTI, BstMBI, DpnII ↓↓↓GATC NZYBuffer C 80 80 100 100 NZYBuffer U * (MB233) Kzo9I, MboI, NdeII, Sau3AI EcoRI G↓↓↓AATTC NZYBuffer A 100 (120) (80) 100 NZYBuffer U * FunII (MB067) EcoRV GAT ↓↓↓ATC NZYBuffer A 100 0 25-50 100 NZYBuffer U * Eco32I (MB068) HindIII A↓↓↓AGCTT NZYBuffer B <20 100 (100) 100 NZYBuffer U * No None (MB070) HinfI G↓↓↓ANTC NZYBuffer C (90) (90) 100 100 NZYBuffer U * None (MB239) HpaI GTT ↓↓↓AAC NZYBuffer B 20 100 (80) 100 NZYBuffer U * KspAI (MB071) KpnI GGTAC ↓↓↓C NZYBuffer C 50 (80) 100 100 NZYBuffer U * No Acc65I ♦♦♦, Asp718I ♦♦♦ (MB072) BfuCI, Bsp143I, BstENII, BstMBI, DpnII, MboI ↓↓↓GATC NZYBuffer C 100 100 100 100 NZYBuffer U * (MB241) Kzo9II, NdeII, BstKTI ♦♦♦ MluI A↓↓↓CGCGT NZYBuffer A 100 100 100 100 NZYBuffer U * None (MB243) Buffer Enzyme Activity in NZYTech buffers (%) Buffer for Restriction site Methylation Reaction Enzyme provided for double Isoschizomers (5’ →→→3’) A (green) B (blue) C (red) U (white) sensitive Conditions 100% activity digestion NcoI C↓↓↓CATGG NZYBuffer B 20 60 + (60) 100 NZYBuffer U * No Bsp19I (MB066) NdeI CA ↓↓↓TATG NZYBuffer A 100 100 80 100 NZYBuffer U * No FauNDI (MB069) NheI G↓↓↓CTAGC NZYBuffer C <20 <20 100 100 NZYBuffer U * AsuNHI, BmtI ♦♦♦, BspOI ♦♦♦ (MB063) PstI CTGCA ↓↓↓G NZYBuffer A 100 80 (20) 100 NZYBuffer U * No BspMAI (MB073) RsaI GT ↓↓↓AC NZYBuffer C 80 100 100 100 NZYBuffer U * AfaI, Csp6I ♦♦♦, CviQI ♦♦♦, RsaNI ♦♦♦ (MB245) SalI G↓↓↓TCGAC NZYBuffer A 100 (20) <20 100 NZYBuffer U * None (MB077) ♦ ♦ ♦ ♦ SmaI CCC ↓↓↓GGG NZYBuffer C 0 <20 100 100 NZYBuffer U * Cfr9I ♦♦, PspAI ♦♦, XmaI ♦♦, XmaCI ♦♦ (MB075) SpeI A↓↓↓CTAGT NZYBuffer C 100 100 100 100 NZYBuffer U * No AhlI, BcuI (MB247) TaqI T↓↓↓CGA NZYBuffer C 80 80 100 100 NZYBuffer U * None (MB235) XbaI T↓↓↓CTAGA NZYBuffer U 50 80 50 100 NZYBuffer U None (MB308) XhoI C↓↓↓TCGAG NZYBuffer A 100 160 60 100 NZYBuffer U * PaeR7I, Sfr274I, SlaI (MB074) Legend: () Weak star activity is detected + 100% activity is obtained by addition of 0.01% BSA (final concentration) * Buffer not provided. Sold separately – Cat. No. MB110 Letter code: N = G, C, A or T R = A or G Y = C or T ↓↓↓ cleavage site ♦♦♦ Neoschizomer (recognizes the same sequence but has a different cleavage site) DNA cleavage blocked or impaired by CpG, dcm or dam methylation DNA cleavage blocked or impaired by overlapping CpG, dcm or dam methylation DNA cleavage potentially inhibited by CpG, dcm or dam methylation Enzyme reaction conditions (temperature and time) Thermal inactivation conditions (temperature and time) .