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文献和实验the melted agarose up/down to get a uniform mixture. Total volume of the gel slice should be ~40 µl. Can freeze the gel slice to store. Ligation Add 2 µl 10X ligase buffer to a tube (the commercial NEB stuff is fine). Add 2 µl water. Melt your gel slices
. Transfer the supernatant to a new 1.5 ml microcentrifuge tube and ethanol precipitate. 14. Resuspend the dried DNA pellet in 40 ul of 10:0.1 TE buffer and assay for concentration by agarose gel electrophoresis. C. Miniprep double-stranded
is between 0.1 and 1 μg for genomic DNA for a total reaction mixture of 100 μl. Larger template DNA amounts usually increase the yield of non-specific PCR products. Primers. (1) PCR primers should be 10-24 nucleotides in length. (2) The GC content should be 40%-60
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