For the PCR, 50 ng genomic DNA as the template, 0.2 mmol/L
dNTPs (Fermentas), 1.5 mmol/L MgCl2, 10 pmol each of the primers,
1.25 units of Taq DNA polymerase (5 U/mL, Fermentas) and 5 mL
Mg2þ free Taq DNA polymerase buffer were combined in a 50 mL
final reaction volume. PCR amplification conditions included 300 s
initial denaturation step at 94 C, 40 amplification cycles (60 s
denaturation at 94 C, 60 s annealing at 46 C, 60 s elongation at
72 C), and 6000 s final extension at 72 C. S. thermophilus CCM
4757 DNA was used as template for positive control reactions.
For the PCR, 50 ng genomic DNA as the template, 0.2 mmol/LdNTPs (Fermentas), 1.5 mmol/L MgCl2, 10 pmol each of the primers,1.25 units of Taq DNA polymerase (5 U/mL, Fermentas) and 5 mLMg2þ free Taq DNA polymerase buffer were combined in a 50 mLfinal reaction volume. PCR amplification conditions included 300 sinitial denaturation step at 94 C, 40 amplification cycles (60 sdenaturation at 94 C, 60 s annealing at 46 C, 60 s elongation at72 C), and 6000 s final extension at 72 C. S. thermophilus CCM4757 DNA was used as template for positive control reactions.
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