1 Department of Clinical Biochemistry, Sher-I-Kashmir Institute of Medical Sciences, Srinagar, Kashmir, India.
2 Department of Immunology and Molecular Medicine, Sher-I-Kashmir Institute of Medical Sciences, Srinagar, Kashmir, India.
3 Department of Biotechnology, University of Kashmir, Srinagar, Kashmir, India.
4 Department of Biosciences, Jamia Millia Islamia, New Delhi, India.
* Corresponding author: Dr. Mushtaq A. Siddiqi, Professor & Chief, Department of Immunology and Molecular Medicine, Sher-i-Kashmir Institute of Medical Sciences, Soura, Srinagar, Kashmir, India 190011.
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The main aim of this study was to compare the different methods of DNA extraction from snap-frozen tissues (SFTs). The samples were submitted to a six different DNA extraction methods -four manual and two using commercial DNA isolation kits. All the retrieval methods were compared with each other for DNA quality and yield. DNA was qualified and quantified by spectrophotometer analysis, electrophoresis, and amplification by PCR. The 237 bp and 115 bp amplicons of p53 gene and K-Ras gene respectively, were amplified equally from DNA extracted by all tested methods and in all cases. According to our results, the extraction method using any salt (4M) proved to be simple and suitable and time saving for obtaining DNA of good quality, which can be easily amplified by PCR. However, the DNA extracted by manual methods had low purity as compared to DNA isolated using kit but the DNA yield by manual methods was quite high than that of the kits.
