Assessment of extracted DNA from whole blood and buffy coat in healthy individuals

Abstract

Deoxyribonucleic acids (DNA) are the genetic material in all organisms. Extraction of DNA is the initial gait for the field of molecular biotechnology and biomedical research to advance understanding of human illnesses from various diseases and provide practical solutions for diagnosis and treatment. Routine diagnostic techniques in medicine have been greatly enhanced by molecular biology. Due to research on deoxyribose nucleic acid, it frequently finds use in medicine. Efficient isolation of DNA from a sample is the basis for successful forensic DNA profiling. DNA can be extracted from variety of samples such as a whole blood, buffy coat, hair, skin, tissue, urine, buccal swab, saliva etc. Numerous molecular biologic applications require genomic DNA (gDNA) in high quality and in adequate quantity. We extracted genomic DNA from buffy coat and whole blood samples and compared the results of the product obtained in terms of quantity (concentration of DNA extracted and DNA obtained per ml of blood used) and quality (260nm/280nm ratio) of the obtained yield. The aim of this study was to extract DNA from whole blood and buffy coat and compare quantity or Concentration (μg/ml) and purity (260nm/280nm) of extracted DNA. The present study was a hospital- based comparative and experimental study. The study was conducted over a period of 1 year on 50 samples. The venous blood sample is taken from the healthy individuals who are willing to donate blood in blood bank. The 5 ml of blood sample taken by venipuncture from healthy individuals were equally dividing into two anticoagulant EDTA tubes by equal volume. The first tube was use as it is, as a whole blood sample and second tube were centrifuged to obtained three phase separation and prepare buffy coat. From both of the samples extraction of DNA is carried out by follow the protocols of modified salting out method. The integrity and purity of the extracted DNA are assessed by Thermo Scientific Nano Drop 2000Spectrophotometers. Our results show thatMean±SD of concentration of DNA from whole blood and buffy coat was 37.49±25.31 μg/ml and 101.83±31.22 μg/ml (P < 0.0001). The Mean±SD of purity (260nm/280nm) of DNA from whole blood and buffy coat was 1.66±0.16 and 1.74±0.15 (p≤0.05). The both p value was found to be statistically significant.The present study concluded that DNA extracted from Buffy coat had purity as well as mean concentration significantly higher when compared to DNA extracted from whole blood.