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文献和实验Troubleshooting for PCR and multiplex PCR
of these ingredients in the PCR mix are: 50 mM KCl; 10 mM Tris-HCl; 1.5 mM MgCl2).QUESTIONS and SOLUTIONS1. I get (many) longer unspecific products. What can I do? Decrease annealing time Increase annealing temperature Decrease extension time Decrease extension
-crack to help liberate the DNA. Check that the solution actually freezes. Overlay with a drop of mineral oil and incubate at 60°C for 60 minutes, followed by 95°C for 15 minutes. Cool to 4°C. Pipette 22.5 l of PCR master mix
can be amplified. Aim PCR amplification of the P2X2-specific fragment will be performed using the following primers which introduces restriction sites in order to facilitate manipulation of the amplified DNA product at a later stage
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