5X Long PCR Buffer¶
Contributed by Ian Chin-Sang, Queens University, ON, Canada
5X Long PCR Buffer (store in 1ml aliquots in -20C).
This buffer seems to work well with Worm Genomic DNA using Taq/Pfu (3:1) mixture.
425 mM KOAc 125 mM Tricine pH 8.7 (adjust pH of Tricine stock solution with KOH) 40% glycerol 5% DMSO 1.2 mM Mg(OAc)2
- Combine ingredients in a suitable container. For 10ml 5X long PCR buffer use:
- 4.25 ml 1M KOAc
- 1.25 ml 1M Tricine, pH8.7 @ 25 degrees C (with KOH)
- 4.00 ml glycerol
- 0.50 ml DMSO
- 120 ul 500 mM Mg(OAc)2
- Store in 1ml aliquots at -20’C until required.
This method is based, with permission, on an original protocol available here.