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Table 1 Composition of PCR-RFLP reaction mix and PCR conditions

From: Sociodemographic and genetic determinants of nonalcoholic fatty liver disease in type 2 diabetes mellitus patients

PCR reaction mix
 Taq buffer (10×) 2.5 μL
 MgCl2 (25 mM) 2.5 μL
 dNTP mix (20 mM) 2.5 μL
 Forward primer (10 μM) 1.5 μL
 Reverse primer (10 μM) 1.5 μL
 Taq DNA polymerase (5 units per μL) 0.5 μL
 Water 12.0 μL
 Genomic DNA 2.0 μL
 Total volume 25.0 μL
PCR conditions
 Denaturation initial 94 °C for 2 min 01 cycle
 Denaturation subsequent cycles each 94 °C for 30 s 35 cycles
 Annealing 66 °C for 30 s
 Elongation initial 72 °C for 30 s
 Elongation final 72 °C for 5 min 01 cycle
RFLP reaction mix
 PCR product 10.0 μL
 Nuclease-free water 18.0 μL
 Tango buffer 10× 2.0 μL
 BtsCI enzyme 1.0 μL
 Total volume 31.0 μL