• E. Sreedevi
• T. Pullaiah

A protocol was developed for DNA isolation and PCR conditions of Alstonia scholaris (L). R. Br., a medicinal plant of Apocyanaceae for conservation concern containing high levels of alkaloids and secondary metabolites. It involves extraction employing liquid nitrogen for grinding and the powdered tissue was transferred to extraction buffer. The quality of DNA was further tested by agarose gel electrophoresis. The yield of DNA ranged from 1049.1 and 877.2 ng (50 μl final volumes) from 3 gm of mature leaf tissues and the purity (ratio) was between 1.7-1.8 indicating minimal levels of contaminating metabolites. This is ideal for isolation of DNA and the DNA isolated was used for randomly amplified polymorphic DNA (RAPD) analysis. RAPD protocol was based on the use of higher concentration of MgCl2 (2.5 µl), lower concentrations of primer (1.0 μl) and Taq polymerase (0.2 units), 2.0 μl of template DNA and an annealing temperature of 35°C, resulted optimal amplification. Reproducible amplifiable products were observed in all PCR reactions. Thus the results indicate that the protocol was suitable to plant species belonging to different genera with high levels of polysaccharides and polyphenolics and for processing large number of samples for genomic analysis, mapping and next generation sequencing.

PCR, CTAB method, DNA Isolation, RAPD Analysis, Genetic Diversity

"DNA Extraction and Assessment of genetic variability through RAPD analysis for Molecular Studies in Alstonia scholaris (L.) R. Br.", International Journal of Emerging Technologies and Innovative Research (www.jetir.org), ISSN:2349-5162, Vol.6, Issue 6, page no.480-487, June-2019, Available :http://www.jetir.org/papers/JETIR1908229.pdf

"DNA Extraction and Assessment of genetic variability through RAPD analysis for Molecular Studies in Alstonia scholaris (L.) R. Br.", International Journal of Emerging Technologies and Innovative Research (www.jetir.org | UGC and issn Approved), ISSN:2349-5162, Vol.6, Issue 6, page no. pp480-487, June-2019, Available at : http://www.jetir.org/papers/JETIR1908229.pdf