Laxmi Kant Pandey, Satya Prakash Tantuway, Yasmin Bano, Nitin Swamy, Ranjan Singh, Sundaram Bhuvaneswari
There is an increasing international market for medicinal plants, which are used both for herbal medicine and for pharmaceutical products. For rapid and accurate identification of plants DNA barcoding is most authentic tool. The aim of this study is first optimize a method for the genomic DNA isolation from non-identified, carbohydrate rich, plant leaf tissues. Than do the DNA barcoding by using the ITS 2 as a potential molecular marker. In this study we first optimize a method for the genomic DNA isolation from non identified, carbohydrate rich, plant leaf tissues. For the optimization of DNA isolation method different combination and permutation were done at different stages of the DNA isolation. Further, the genomic DNA was utilized as a template for the amplification and sequencing of ITS2 region. BLAST search was performed against GenBank database by using annoted ITS2 region as sample query. We found very high yield around 123 µg per gram of leaf samples and the ratio at 260/280 nm found to be 1.82. Hence the yield as well as quality of the DNA was very good. Two PCR products of 419 and 390 bp were amplified by using ITS2F1, ITS2F2 as forward and ITS2R as reverse primers respectively. In annotation out of 419, 228 bp sequences were annotated as ITS2 for this plant in blast search it shows close resemblance to the plants of plantaginaceae in the order lamiales. In herbarium identification the plant was identified as Anisochilus carnosus (Lf) which belongs to Lamiaceae family in the order lamiales.
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