Method for direct amplification from crude nucleic acid samples
View Patent ↗The present teachings relate to improved methods, kits, and reaction mixtures for amplifying nucleic acids. In some embodiments a novel direct buffer formulation is provided which allows for the direct amplification of the nucleic acids in a crude sample with minimal sample purification.
1. A method comprising providing a paper comprising a lysis reagent spotting blood or collecting buccal cells on the paper comprising the lysis reagent placing the blood on paper or the buccal cells on paper in a direct buffer to form a nucleic acid containing solution and performing a PCR on the nucleic acid containing solution, wherein the direct buffer comprises 0.2%-0.9% polysorbate, 3%-8% glycerol, and 1000-3000 ug/ml BSA.
2. The method according to claim 1 , wherein the direct buffer further comprises, 10-50 mM Tris-HCl (pH 8.3), 30-80 mM KCl, 1.4-2.4 mM MgCl 2 , 0.01%-0.04% Sodium azide, 100-350 uM of each dNTPs, and 0.10-0.35 U/ul of DNA polymerase.
3. The method according to claim 1 , wherein polysorbate is polysorbate 20.
4. The method of claim 1 , wherein the nucleic acid containing solution further comprises a plurality of primer pairs, wherein the plurality of primer pairs flank a plurality of short tandem repeat loci.
5. The method of claim 1 , wherein the paper is Fast Technology for Analysis of nucleic acids (FTA) paper.