IP Library › Granted Patent US 8,460,934
Granted Patent B2
US 8,460,934 · App. 13/438,436 · Granted Jun 11, 2013

Method for direct amplification from crude nucleic acid samples

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Quick Facts
Patent No.
US 8,460,934
App. No.
13/438,436
Granted
Jun 11, 2013
Kind
B2
Abstract

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.

Claims (5)

1. A kit comprising a plurality of primer pairs and a direct buffer, wherein the direct buffer comprises 0.2%-0.9% polysorbate, 3%-8% glycerol, and 1000-3000 ug/ml BSA.

2. The kit of 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, 1200-3000 ug/ml BSA, and 0.10-0.35 U/ul of DNA polymerase.

3. The kit of claim 2 , wherein the polysorbate is polysorbate 20.

4. A reaction mixture comprising a direct buffer and a plurality of primer pairs, wherein each primer pair flanks a genomic locus containing a short tandem repeat (STR), and wherein the direct buffer 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, 3%-8% glycerol, 100-350 uM of each dNTPs, 0.2%-0.9% polysorbate 20, 1200-3000 ug/ml BSA, and 0.10-0.35 U/ul of DNA polymerase.

5. The kit of claim 1 , wherein each primer pair flanks a genomic locus containing a short tandem repeat.