Abstract
Conclusion: PPAR-α gene has important role in
the homeostasis of lipid profile in the body.
Upregulation or increased expression of this gene
leads to reduction in lipid profile, while down
regulation or suppression of this gene leads to
deranged lipid levels. (Rawal Med J 202;45:54-
57).
Results: Primers were optimized using
conventional PCR. A single band at 224 base pair
represented the amplification of PPAR-α gene.
The PCR protocol was optimized for PPAR-α
gene.
Methodology: This experimental study was
carried out at Center for research in experimental
and applied medicine (CREAM) Department of
Biochemistry and Molecular Biology, Army
Medical College, Rawalpindi from December 20,
2016 to June 8, 2017. The study comprised of
three groups; Group one included subjects who
were healthy, group two was comprised of diabetic
without dyslipidemia and group three included
diabetics with dyslipidemia. Blood was collected
and Primers specific to PPAR-α were designed
on the basis of sequence available on website
Primer BLAST. RNA was extracted with in first six
hours of sample collection using Gene JET RNA
purification kit (Thermo Scientific). For first strand
cDNA synthesis Revert Aid Premium was utilized.
Polymerase chain reaction was used for the
isolation, amplification or detection of specific
nucleotide sequences in genomic DNA. PCR
products were visualized by Agarose gel
electrophoresis. After Real time PCR, molecular
analysis was quantified by comparative CT
Method (ΔΔ Ct method). Positive value means
down regulation and negative value means
upregulation.
