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文献和实验CCMB buffer to make chemical competent cell
and kanamycin appear to do fine with 1 hour growth Plate 20 μl on AMP plates using 3.5 mm glass beads Transformation efficiency is 15 x colony count x 105 There is anecdotal evidence that refreezing partially used cell aliquots dramatically reduces
Diatomaceous Earth-based Midi-prep
as above for 11-14 hours. Harvest cells by transferring into a 50 ml sterile conical tube and spinning at 3000 rpm in GPR tabletop centrifuge for 5 min. Decant supernatant and freeze cell pellet at -70oC for at least 1 hour. Resuspend defrosted cells in 2 ml
Diatomaceous Earth-based Midi-prep
by transferring into a 50 ml sterile conical tube and spinning at 3000 rpm in GPR tabletop centrifuge for 5 min. Decant supernatant and freeze cell pellet at -70oC for at least 1 hour. Resuspend defrosted cells in 2 ml GET/lysozyme (2 mg/ml). Add 4 ml
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