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Effect of Long Term Storage of Buccal Swab on Pcr of Hvi Segment of Mitochondrial DNA at Freezing Point

Solomon AY, Danborno SB, Timbuak J and Nachana ND

Effect of Long Term Storage of Buccal Swab on Pcr of Hvi Segment of Mitochondrial DNA at Freezing Point

Amplification of human mitochondrial DNA have been widely used in population genetics in human evolutionary and molecular anthropology studies. Mitochondrial DNA HVSI was shown to be a suitable tool in genetic analysis due to the unique properties of the mitochondrial DNA, such as the lack of recombination, maternal mode of inheritance, rapid evolutionary rate and high population specific polymorphisms. In this work, the effect of long term storage of buccal swab cells at freezing point was assessed and the DNA yield quantity was determined. The aim and objectives of this study was to determine the integrity of the DNA of the HVI segment obtained from buccal swab stored at long term at freezing point and to gel electrophorose in order to determine the yield and purity of the HVSI of mitochondrial DNA. Materials used were Accruprep genomic extraction kit, sterile buccal swab, micro pipette, micro pipette tips, racks, vortex, centrifuge, binding column, Eppendorf tubes, primers, gel box, PCR machine, TBE buffer, incubator, photo spectrometer.Samples (Buccal swabs) were collected from 15 individuals and separated into 3 groups, group A was processed immediately after collection, group B and C were stored at freezing point for 15 and 30 days respectively and then processed.Genomic DNA was extracted from buccal epithelial cells according to the protocol of the manufacturer (Accrupep genomic extraction kit). The effect of long term storage of buccal cell swab on the suitability of the samples for PCR amplification efficiently amplifiedat 360bp fragments of HV I segment All examined PCR products, were successfully purified regardless of the sample storage time and condition.The purity of genomic DNA was estimated by OD260/OD280 ratio. The results suggest that DNA samples stored at freezing point for a long period of time shows no significant difference in the amount, purity and yield of Mitochondrial DNA.

Key Words: PCR, Buccal swabs, Hypervariable I segment, Mitochondrial DNA

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