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) | |
在培养板中用牙签挑取菌落后悬浮于100 μl dH2O中。取10 μl作为PCR反应的模板: | ||
PCR反应条件 | ||
98℃ 5 sec. | 25 cycles(约60 min) | |
55℃ 5 sec. | ||
72℃ 90 sec. | ||
(TaKaRa Thermal Cycler PERSONAL, Fast mode) | ||
【结果】 | ||