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An Efficient Procedure for DNA Isolation and Profiling of the Hyper Variable MtDNA Sequences



Author(s)

Nazia Akbar, Habib Ahmad, Muhammad Shahid Nadeem, Nasir Ali and Muhammad Saadiq

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DOI:10.17265/1934-7391/2015.11.004

Present study describes the results of an efficient protocol for the isolation of good quality DNA from human saliva. The protocol includes collection of saliva in sterile specimen tubes, followed by the cell lysis. After formation of cell lysate, proteins wereextracted by phenol chloroform treatment for purification of DNA. The purified DNA was precipitated by adding equal volume of isopropanol to the treated supernatant. After isolation DNA pellet was washed with 70% ethanol, air-dried and was suspended in 30µL of double distilled water. Best quality of DNA was extracted from the saliva samples and the PCR product was amplified for hyper-variable regions (HV1& HV2) of the mitochondrial DNA. The genes were cleaned with GeneAll gel elution kit (Gel SV) (Cat. No. 102-10) and sequenced accordingly. The DNA isolation protocol presented here is recommended for the isolation, best quality and yield of DNA from the human saliva.

Human saliva, DNA isolation, mtDNA sequences, forensic study