High efficiency PCR

Total volume 5...10 µL:

• 45 mM Tris-HCl (pH 8.8)
• 11 mM ammonium sulfate
• 4.5 mM MgCl
• 6.7 mM beta-mercaptoethanol
• 4.4 µM EDTA (pH 8.0)
• 1 mM each dNTP
• 113 µg/mL BSA
• 2 pmol of each primer
• 0.5 U Taq-polymerase

Buffer can be prepared as 10x stock and stored at -20°C.

Due to low volumes you can use short denaturing, annealing and elongation times (typically 15...30 sec).

Ref.:

Jeffreys AJ, Neumann R, Wilson V.: Repeat unit sequence variation in minisatellites: a novel source of DNA polymorphism for studying variation and mutation by single molecule analysis. Cell. 1990 Feb 9;60(3):473-85. [PubMed]

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