Manuscript Title:

A MODIFIED CTAB METHOD FOR DNA EXTRACTION OF OILY PLANTS OF BRASSICACEAE FAMILY

Author:

MAIDA ASLAM, FATMA HUSSAIN, NISAR AHMED, AMER JAMIL

DOI Number:

DOI:10.5281/zenodo.10123210

Published : 2023-11-10

About the author(s)

1. MAIDA ASLAM - Molecular Biochemistry Lab, Department of Biochemistry, University of Agriculture Faisalabad, Pakistan.
2. FATMA HUSSAIN - Department of Biochemistry, University of Agriculture Faisalabad, Pakistan.
3. NISAR AHMED - Centre of Agricultural Biochemistry and Biotechnology (CABB), University of Agriculture Faisalabad, Pakistan.
4. AMER JAMIL - Molecular Biochemistry Lab, Department of Biochemistry, University of Agriculture Faisalabad, Pakistan.

Full Text : PDF

Abstract

Extraction of DNA from secondary metabolite enriched medicinal plants has always been tricky. Numerous methods for DNA extraction has been presented but those either gives poor yield or employed to least chemical heterogenetic plants species with use of costly and hazardous chemicals and conditions that become worse for oily plants. To our knowledge only a few protocols have been reported for DNA extraction from oily plants that not only are laborious but also involve very complex reagents. Here we report simple DNA extraction strategy comparable to costly commercial DNA extraction kits which is equally potent for oily and non-oily plants. High-molecular weight DNA was extracted without the use of liquid nitrogen and RNase A treatment that markedly reduce the extraction cost. Quantity of the DNA extracted from species with lower nuclear DNA content was in the range of 250-380 μg/mL with A260/A280 absorbance ratio 1.67- 1.87 that confirmed purity of the recollected DNA samples. Quality of the extracted DNA was also evaluated by restriction analysis and PCR amplification. This protocol gives DNA that is suitable for molecular biology techniques sensitive to contaminants.


Keywords

Nuclear DNA, Oily Plants, Brassicacea, CTAB (Cetyl Trimethyl Ammonium Bromide), A260/A280 Absorbance, RNaseA.