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江苏天净沙
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文献和实验Screening lambda cDNA or genomic libraries
- If you store it frozen,take the alpha 32P-dATP out of the freezer to thaw. - Put 50 ng DNA to be labelled in a total volume of 18.9 µl H20.For labelling fragments from a restriction digest: I prefer not to do the labelling in the low melt agarose
糖凝胶分离的范围 试剂: (1)1 X TAE电泳缓冲液:45mmol/Ltris-硼酸,1mmol/LEDTA,pH8.0 (2)凝胶加样缓冲液:0.25%溴酚蓝,40%蔗糖水溶液。 (3)λ-DNA和提取的DNA λ-DNA是λ噬菌体的基因组DNA,全长50kb。用HindIII或(和)BamHI酶切得到的片段被广泛用于DNA电泳的分子量标准。 (4)分子量标准(&lambda
Recombineering/Lambda red-mediated gene replacement
in -80°C freezer. Transform strain with pKD46 and grow on LB-amp plate at 30°C Prepare four tubes with 0, 1 ng, 10 ng, and 100 ng pKD46 plasmid DNA Add 100 μL of competent cell mix to each tube
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