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DNA的重组  (一)DNA的酶切与连接  (1)酶切反应  同质粒DNA的鉴定,只不过是质粒DNA换为载体DNA。若大量酶切,则成比例增加。  (2)加2倍体积的预冷无水乙醇和1/10体积的3mol/l Na
An inexpensive alternative to glassmilk for DNA purification...
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An inexpensive alternative to glassmilk for DNA purificationPreparation of Silica1. Suspend 5 g of silica (Sigma, S-5631) in 50 ml of PBS. 2. Allow the silica
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Electroelution of agarose fragmentsElectroelution buffer1 M Tris, pH 7.5 12.0 mls0.5 M EDTA 0.24 mls1 M NaCl 3.0 mlsqs to 600 mls dH2OAcetate cushion 3 M NaAce
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Cool growth competent E . coliThis protocol is very effective and simple. The fact that the cells are grown at low temperature greatly increase the OD range du
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Electroporation of P. aeruginosa Preparation of Electrocompetent Cells Inoculate a 5-ml of L-broth with cells of the P. aeruginosa strain to be electrotransfor
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Preparation of Electrocompetent Cells and Electroporation of Plasmid DNA Inoculate 5-ml L-broth with a single colony of E. coli. Incubate 5 hours to overnight
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FROZEN COMPETENT E. COLI CELLS(Inoue et al., 1990 Gene 96:23) Inoculate a 5ml overnight of E.coli in LB+20 mM MgSO4. Next morning, inoculate 250 ml LB+20 mM Mg
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Inoue, H., H. Nojima, and H. Okayama. 1990. High efficiency transformation of Escherichia coli with plasmids. Gene 96:23-28. Citation Abstract Procedure1. Inoc
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