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文献和实验in a low speed centrifuge at 2000 rpm for 5 minutes. If I ligate 100 ng of blunted dephosphorylated vector to 50 ng of a blunt 500 bp PCR in a 30 µl reaction, I'll usually transform 5 ul into cells and plate out the whole tranformation, I normally
DNA Recovery With Low Melt Agarose
Recovery of DNA from Low Melting Point Agarose Gels 1.Run digestion products on 0.7% LMP agarose gel in 1X TBE (it's nice to have at least 1ug of the fragment you want). LMP agarose is fragile; pour gel with EtBr 2.Let solidify in cold room
Preparation of Mycoplasma chromosomal DNA in low melting agarose
1. Spin 1.5ml fresh cells for 3 minutes. 2.Wash the pellet once with TNE buffer. 3. Suspend the pellet well in 20 ul TNE buffer. 4. Add 2 ul proteinase K ( stock 10 ug/ul in water, -20 C). 5. Add 20 ul low melting agarose (1.6% in TNE buffer
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