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- 保存条件:
20℃
- 保质期:
12个月
- 库存:
大量
- 供应商:
钰博生物
- 规格:
20 rxns
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文献和实验TOP10 chemically competent cells
Overview This protocol is a variant of the Hanahan protocol [1] using CCMB80 buffer for DH10B, TOP10 and MachI strains. It builds on Example 2 of the Bloom05 patent as well. This protocol has been tested on TOP10, MachI and BL21(DE3) cells
takes ~2.5 hours) -Spin cells in two 50ml Falcon or Corning disposable centrifuge tubes at 2.5 Krpm for 5min at 4℃. (Just enough to pellet the cells). -Drain cells and resuspend in 20-40mls per 100mls starting culture of TfbI. You can be quite
Frozen competent E. coli cells 【Carnegie Institution of Washington】
Inoculate a 5ml overnight of E.coli in LB+20 mM MgSO4. Next morning,inoculate 250 ml LB+20 mM Mg++ in a 2L flask with about 2ml overnight culture.Grow at room temp (23℃)with good aeration (250rpm)to an A600 of 0.4-0.6.Temp is important
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