挑取菌落&PCR | ||
【方法】 | ||
plasmid: | 8 kb insert in pUC18 | |
宿主: | JM109菌株 | |
primer: | M13 primer M4 and RV | |
polymerase: | Z-Taq(2.5 U/μl) | |
在培養(yǎng)板中用牙簽挑取菌落后懸浮于100 μl dH2O中。取10 μl作為PCR反應(yīng)的模板: | ||
PCR反應(yīng)條件 | ||
98℃ 5 sec. | 25 cycles(約60 min) | |
55℃ 5 sec. | ||
72℃ 90 sec. | ||
(TaKaRa Thermal Cycler PERSONAL, Fast mode) | ||
【結(jié)果】 | ||