The Biobrio 9(1 & 2), 2022

Optimization of genomic DNA isolation method from mature leaves of Dalbergia latifolia

Aditya Kumar, Praveen Paul Horo, Shilpi Kumari and Subhash Prasad

ABSTRACT:

Germplasm conservation of Dalbergia latifolia is very important as it is listed in vulnerable category of IUCN Red list. To reduce the efforts, cost, land and labour in conservation of germplasm, it is needed to assess genetic diversity present in the species. The success of genetic improvement program also depends on genetic diversity present in the breeding material. The molecular markers are widely utilized for the assessment of genetic diversity these days. High quality and quantity of genomic DNA is prerequisite for utilizing molecular markers. Hence, the protocol for the isolation of genomic DNA of black shisham (D. latifolia) has been optimised under the present study. The optimized protocol yielded 500-9000 ng/μl genomic DNA with an average of 2500 ng/μl DNA from 89 samples of Dalbergia latifolia. A very low level of RNA, protein, phenolics and polysaccharide contaminants were recorded (A260/A280 ratio ranges from 1.56-2.06) from the isolated DNA. The optimized protocol was yielded genomic DNA of good quality and quantity from both, juvenile leaf and matured leaf samples so, the protocol can be utilized for isolation of high-quality DNA for genetic improvement programs and genetic diversity assessment.

Keywords:    

Vulnerable, Dalbergia latifolia, molecular markers, genomic DNA, genetic improvement

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