An Improved PCR-RFLP Assay for the Detection of a Polymorphism of PLIN1 Gene
Background:
In recent research, it has been shown that there have been variants of rs894160 within the PLINI gene which have been associated with obesity, type 2 diabetes, and other diseases. But the isoschizomers such as the Mn1I enzyme required for the detection of this polymorphism are expensive.
Methods:
The study used an improved PCR-RFLP method with mismatched base for detection of the single nucleotide polymorphism rs894160.
Results:
After detecting 550 Chinese Han individuals, the genotype frequencies were 26.0% for AA, 50.0% for AG, and 24.0% for GG. The allelic frequencies were 51.0% for A and 49.0% for G. The PCR results were confirmed by DNA sequencing. The chi2 test showed the genotype and allele frequencies of PLIN1 do not deviate from Hardy-Weinberg equilibrium, and the sequences of amplified products were consistent with the one published in Genbank with the exception of mismatched base.
Conclusions:
Based on the PCR with mismatched primers we designed, the PLIN1 polymorphisms could be identified effectively.
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