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文献和实验Mouse p27 PCR Using Gitschier Buffer
Mutant allele: Neo-1 primer (CCTTCTATCGCCTTCTTG), plus mgK-3 primer (TGGAACCCTGTGCCATCTCTAT) produce a 0.5kB PCR product. Wildtype allele: mgK-3 (above) plus mcK-5 (GAGCAGACGCCCAAGAAGC) produce ~1kB product. Stock Solutions
Adding 3A overhang to a PCR product
1 PCR Buffer with Mg (10x) 1x (1.5 mM MgCl2) 5 Taq DNA Polymerase (5 U/μl) 1U 0.2 ddH2O
PCR product purification-Molecular Biology
specified in the kit2) Qiaquick is the gel kit, right? So you actually have to cut out your product. How intense was your band of interest? Maybe your PCR wasn't particularly good3) Check the pH of your elution buffer, or (especially!) water, DNA
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