E.coli BMH 71-18 mutS Competent Cells
Contents
1. TakaRa Tel 077 543 6116 Fax 077 543 197 http wWW taKara DiO CO jp 201109Da2. http wwwr takara bio coJp 2 9054 1 2 14 ml BD Code 352059 352057 1 5 ml 3 100 ul DNA 10 ng 4 42 60 5 SOC L broth b broth L broth 10 g Bacto tryptone 5 g Bacto yeast extract 5 g NaCl 1 L water 1NKOH pH7 5 b broth
3. 9054 IdfaR4 E coli BMH 71 18 mutS Competent Cells vV201109Da 1 E coli BMH 71 18 mutS Competent Cells 100 ul X 10 pUC119 plasmid 0 1 ng ubD 10 ulxX 1 SOC 1ml x 10 SOC 2 Tryptone 0 5 Yeast extract 10mM Nadl 2 5mM KO 10mM MgSO4 10mM MgCb2 20mM Glucose Ill DNA DNA TaKaRa Hanahan Ecoli BMH 71 18 mutS Competent Cells mutS DNA Site directed mutagenesis 2z DNA DNA lI 1 1 Ecoli BMH 71 18 mutS Competent Cells
4. 1 ng pUC119 Amp gt 1 Xx 107 colonies ug pUC119 V Genotype E coli BMH 71 18 mutS A lac proAB supE thi 1 mutS215 Tn10 tet F traD36 proAB t lacA lac A M15 VI Cell density 1 2X102bacteria ml http wwwr takara bio coJp 3 9054 VII 80 80 pUC119 VIlI 1 Hanahan D 1983 Mol Biol 166 577 2 Kunkel T A 1985 oc Natl Acad 5 USA 82 488 3 Kunkel T A etal 1987 Methods in Enzymology 154 367 4 Zoller MJ and Smith M 1983 Methods in Enzymology 100 468 5 1986 vol 1 ll 105 6 Hashimoto Gotoh T et g 1995 Gene 152 271 275 IX pUC119 DNA 3319
5. 2 100 ul 14 ml 3 DNA 10 ng 4 30 5 42 45 6 1 2 7 37 S0C 1 ml 8 37 1 160 225 rpm 9 10 37C 9 cm 100 ul 2 M13 DNA 1 1 8 2 3 ml YT soft agar 46 48 200 ul Eco 7 BMH 71 18 mutS Aeoo 0 8 1 0 3 1 2 agar YT 4 10 15 37
6. 5gBacto yeast extract 20 g Bacto tryptone 5 9 MgSO4 7H20 1Lwater 1NKOH pH7 5 1 7 7 YT 8gBacto tryptone 5 g Bacto yeast extract 5 g NaCl 1 L water 1 N NaOH pH7 5 1 59 6 agar 8 YT softagar 0 8 9g Bacto tryptone 0 5 g Bacto yeast extract 0 5 g NaCl 100 ml water 1N NaOH pH7 6 0 6 agar 9 10 Site directed mutagenesis Mutan K Mutan Express Km 11 80 1 IV
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