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Table 4 Polymerase chain reaction (PCR) reagent mixtures for multiplex single base extension assays.

From: Development of a single base extension method to resolve Y chromosome haplogroups in sub-Saharan African populations

PCR reagent YSNP1 Hg-A Hg-B Hg-B2b
  Concentration Concentration Concentration Concentration
DNA 10 ng/μL 10 ng/μL 10 ng/μL 10 ng/μL
Buffer (10×) 1× 0.8× 1× 1×
MgCl2 (25 mM) 2 mM 4 mM 3 mM 3 mM
dNTPs 2.5 mM) 300 μM 200 μM 200 μM 200 μM
Forward primer (10 μM) See Table 1 See Table 1 See Table 1 See Table 1
Reverse primer (10 μM) See Table 1 See Table 1 See Table 1 See Table 1
FastStart Taq (5 U/μL) 1 U 1 U 1 U 1 U
ddH2O Made up to 25 μL Made up to 25 μL Made up to 25 μL Made up to 25 μL
  Hg-E Hg-E1b1a Hg-E1b1b1  
PCR reagent Concentration Concentration Concentration  
DNA 10 ng/μL 10 ng/μL 10 ng/μL  
Buffer (10×) 1.25× 1.25× 1.25×  
MgCl2 (25 mM 3 mM 3 mM 3 mM  
dNTPs (2.5 mM) 200 μM 200 μM 200 μM  
Forward primer (10 μM) See Table 1 See Table 1 See Table 1  
Reverse primer (10 μM) See Table 1 See Table 1 See Table 1  
FastStart Taq (5 U/μL) 1 U 1 U 1 U  
ddH2O Made up to 25 μL Made up to 25 μL Made up to 25 μL  
  1. dNTP, deoxyribonucleotide triphosphate; ddH20, double distilled water
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Investigative Genetics

ISSN: 2041-2223

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