Journal: BMC Medical Genetics
Article Title: A novel multiplex PCR-RFLP method for simultaneous detection of the MTHFR 677 C > T , eNOS +894 G > T and - eNOS -786 T > C variants among Malaysian Malays
doi: 10.1186/1471-2350-13-34
Figure Lengend Snippet: Agarose gel with PCR–RFLP products for analysis of MTHFR 677 C > T and eNOS +894 G > T and eNOS −786 T > C . Lane 1 contains Fermentas O’ GeneRuler® Ultra Low Range DNA Ladder (25 bp - 700 bp); Lanes 2, 3 and 4 contain multiplex PCR-RFLP products digested with 1 U of NEB® BanII restriction enzyme (NEB®, England), 1 U Fermentas Fast Digest® HinfI restriction enzyme (Fermentas, Lithuania) and 1 U Fermentas Fast Digest® MspI restriction enzyme (Fermentas, Lithuania), respectively. The band sizes were 371 bp, 248 bp, 178 bp, 130 bp and 118 bp for all lanes. This indicates that the sample is a TT genotype for MTHFR 677 C > T, GG genotype for eNOS +894 G > T , and TT genotype for eNOS −786 T > C.
Article Snippet: In these tubes, the RFLP mixture consisted of 8.5 μL of doubly distilled water, 0.1-0.5 ng of fresh PCR product, 1 X Fermentas Fast Digest® Buffer (Fermentas, Lithuania), 1 U Fermentas Fast Digest® HinfI restriction enzyme (Fermentas, Lithuania) and 1 U Fermentas Fast Digest® MspI restriction enzyme (Fermentas, Lithuania).
Techniques: Agarose Gel Electrophoresis, Multiplex Assay