Mini-scale Genomic DNA Extraction from Cotton |
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Authors: | Chaudhry Bushra Yasmin Afshan Husnain Tayyib Riazuddin S. |
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Affiliation: | (1) National Centre of Excellence in Molecular Biology, University of the Punjab, Canal Bank Road, Thokar Niaz Baig, Lahore, 53700, Pakistan |
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Abstract: | Large amounts of polyphenolics in cotton leaves make it difficult to obtain high-quality genomic DNA during extraction. A procedure to isolate nuclear DNA from local cotton leaves (gossypium hirsutum, MNH93, CIM443, FH672) was therefore developed. It consists of rapid isolation of stable nuclei, which hinders covalent interactions with phenolics, followed by DNA extraction. The yield and quality of the resulting DNA is satisfactory and the protocol can be scaled up or down according to sample size. It is suitable for PCR and the restriction enzyme digestion needed for Southern and RFLP analysis. |
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