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[求助] 谁会做GV3101感受态，求教！！！已有1人参与

本人新手，现需要做GV3101感受态，求教程，谢谢！！！

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myprayer: 金币+1, 赠人玫瑰手有余香，分子生物期待你更多精彩。 2015-01-09 19:31:24

One tube of Agrobacterium (GV3101PMP90) was cultured as seed in 20 ml LB medium with Rifampicin (50 mg/l) and Gentamycin (20 mg/l). The seed culture were incubated at 28 °C 180 rpm overnight. Then added the seed culture into 500 ml LB medium with Rifampicin (50 mg/l) and Gentamycin (20 mg/l) and cultured for around 5 h at 28 °C until the OD600 was 0.5-0.7. Before harvesting the cells at 4000 g (Multifuge 3 S-R, Heraeus), 5 min, 4 °C, they were incubated for 10 min on ice. All the following steps were performed on ice as well. Cells were washed by resuspension in 100 ml pre-cooled sterile 150 mM NaCl and spined down at 4000 g, 5 min, 4 °C. The cell pellet was resuspended in 10 ml pre-cooled sterile 20 mM CaCl2. After that, the Agrobacterium were aliquoted (200 ml) into Eppendorf tubes by filling them quickly with a multipette. The aliquots were stored at -80 °C.