Evaluation and Modification of Some Traditional Methods to Extract Genomic DNA in High Quality and Quantity from Different Plant Families | ||
| Egyptian Academic Journal of Biological Sciences. C, Physiology and Molecular Biology | ||
| Volume 16, Issue 2, December 2024, Pages 63-74 PDF (1021.43 K) | ||
| Document Type: Original Article | ||
| DOI: 10.21608/eajbsc.2024.371518 | ||
| Authors | ||
| Mona M. Moghazee1; Shaimaa A. Abdelaziz1; Rehab A. Hasanien1; Rawan S. Zaid1; Rawan Y. Fawzy1; Khaled T. Mohamed2; Ahmad A. Suliman3; Ghada M. Samaha4; Hala M. Zoghly1 | ||
| 1Genetic Department, Faculty of Agriculture, Ain Shams University, P.O. Box 68, Hadayek Shoubra 11241, Qalyubia , Egypt. | ||
| 2Plant Department, Genetics Division, Faculty of Agriculture, Al-Azhar University, Egypt. | ||
| 3Horticultural Crop Technology Department, Agricultural and Biological Institute, National Research Centre, Dokki, Egypt. | ||
| 4Field Crops Research Department, Agricultural and Biological Institute, National Research Centre, Dokki, Egypt. | ||
| Abstract | ||
| Background: Plants are rich in complex compounds, which hinder obtaining intact and pure DNA, especially by traditional manual methods. Extraction of high-quality and quantity genomic DNA is the most important step for all DNA-based biological applications, such as polymerase chain reaction (PCR), cloning, and sequencing. In this study, four DNA extraction modified protocols (CTAB, TNES) and (CTAB, TENS with a solid-phase column) were evaluated to produce highly purifiedand high-quantity DNA from the leaves and seeds of 11 species. They were compared with a kit (QIAGEN) which was used as a positive control. The techniques were evaluated for their capacity to produce DNA devoid of sugars, polyphenols, RNA, and other significant contaminants, as well as their apparent, viscosity, OD260 reading, and appropriateness for PCR-based assays. Results: The most obvious result was that using traditional methods such as CTAB and TENS with a solid-phase column increased the efficiency of the method and gave better results in spectrophotometer readings and polymerase chain reaction. Conclusion: To get results comparable to those from Kit procedures, it is possible to enhance the conventional DNA extraction techniques. To achieve high-quality DNA at the lowest possible cost, this is accomplished by combining several extraction buffers and utilizing a solid phase column. | ||
| Keywords | ||
| DNA extraction protocols; Solid-phase technology; Molecular markers; PCR | ||
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