MvaI (BstNI) (10 U/µL)

Protocol tips

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	To genotype the rs1801320, rs3218536, and rs861539 polymorphisms, 10 μL of each PCR product was digested with either 2 U of MvaI (BstNI) (Thermo Scientific, USA), 1 U of SexAI (New England Biolabs Inc., USA), or 0.5 units of NlaIII (New England Biolabs Inc., USA), respectively, for 16 h at 37°C. The genotypes were determined by running the digested products in 3% agarose gel with ethidium bromide (1 μL/mL) for UV visualization. The products for each genotype of the tested genes are shown in Table 3. Examples of the obtained restriction patterns are presented in Figure 1.

Protocol tips

To genotype the rs1801320, rs3218536, and rs861539 polymorphisms, 10 μL of each PCR product was digested with either 2 U of MvaI (BstNI) (Thermo Scientific, USA), 1 U of SexAI (New England Biolabs Inc., USA), or 0.5 units of NlaIII (New England Biolabs Inc., USA), respectively, for 16 h at 37°C. The genotypes were determined by running the digested products in 3% agarose gel with ethidium bromide (1 μL/mL) for UV visualization. The products for each genotype of the tested genes are shown in Table 3. Examples of the obtained restriction patterns are presented in Figure 1.

Publication protocol

"Single nucleotide polymorphisms (SNPs) of the RAD51 (rs1801320 and rs1801321), RAD51B (rs10483813 and rs3784099), XRCC2 (rs3218536), and XRCC3 (rs861539) genes were analyzed (Table 2).The genotyping of rs1801320, rs3218536, and rs861539 polymorphisms was determined by PCR-RFLP. The primers and PCR conditions for the polymorphic sites of these genes are shown in Table 2. The PCR was run in 10 μL reactions containing 10 ng of genomic DNA, 0.2 mM of each primer, 2.5 mM MgCl2, 1 mM deoxyribonucleotide triphosphates (dNTPs), 3 U HOT FIREPol DNA polymerase, and 1x Solis BioDyne buffer B1. The primers were synthesized by Sigma-Aldrich (USA), and PCR reagents were obtained from Solis BioDyne (Estonia) and Applied Biosystems (USA). Thermal cycling was performed as follows: initial activation at 95°C for 12 min, followed by 30 amplification cycles consisting of denaturation at 95°C for 30 s, annealing at 64°C (rs3218536, rs861539) or 65°C (rs1801320) for 30 s, and extension at 72°C for 1 min, followed by a final extension at 72°C for 10 min.

To genotype the rs1801320, rs3218536, and rs861539 polymorphisms, 10 μL of each PCR product was digested with either 2 U of MvaI (BstNI) (Thermo Scientific, USA), 1 U of SexAI (New England Biolabs Inc., USA), or 0.5 units of NlaIII (New England Biolabs Inc., USA), respectively, for 16 h at 37°C. The genotypes were determined by running the digested products in 3% agarose gel with ethidium bromide (1 μL/mL) for UV visualization. The products for each genotype of the tested genes are shown in Table 3. Examples of the obtained restriction patterns are presented in Figure 1"

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