Comparison of the efficiency of the buffer/detergent extraction and standard salting-out methods for DNA extraction from blood and semen samples

Message:
Abstract:
Genomic DNA extraction with desirable quantity and quality is a basic requirement in molecular biology. Most commercial DNA extraction methods the time-consuming and based on use of enzymes (proteinase K) and toxic organic solvents (phenol-chloroform). Therefore, since these enzymes are expensive and materials are harmful, DNA extraction methods that are void of such disadvantages are required. In this study, buffer/detergent extraction method and salting-out method in different species including blood samples of cattle, sheep, goat and chicken and semen samples of bulls and rams were studied. For DNA extraction and comparison of two methods, 366 blood samples and 26 sperm samples were used. Quantity and quality parameters using Nanodrop and agarose gel (1%) and efficiency PCR (Polymerase Chain Reaction) were evaluated in both methods. Concentration and purity of the extracted DNA were 623.8± 198 µg/mL and 1.87 ± 0.05 respectively in the buffer/detergent method while the concentration and purity of the extracted DNA were 413.1 ±178 µg/mL and 1.82± 0.042 respectively in the salting-out method. Polymerase chain reaction was successful on extracted DNA with either method which proves the absence of inhibitors‌ in the reaction. The electrophoresis results confirmed no breakage in either of the samples. The results of DNA extraction with buffer/detergent method showed that this method was safe, inexpensive and safe, and that the extracted DNA had high quantity and quality; therefore, this method is recommended as an appropriate method for DNA extraction from blood and semen samples in molecular laboratories.
Language:
Persian
Published:
Genetics in the Third Millennium, Volume:8 Issue: 4, 2011
Pages:
2155 to 2161
https://magiran.com/p890971  
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