A simple ethanol wash of the tissue homogenates recovers high-quality genomic DNA from Corchorus species characterized by highly acidic and proteinaceous mucilages
Avijit Kundu1 · Debabrata Sarkar*1 · Amit Bhattacharjee1 · Niladri Topdar1 · Mohit Kumar Sinha2 · Bikash Sinha Mahapatra2
1 Biotechnology Unit, Division of Crop
Improvement, Central Research Institute for Juteand Allied Fibres (CRIJAF),
Barrackpore, Kolkata, West Bengal, India
*Corresponding author: email@example.com
Financial support: Indian Council of Agricultural Research (ICAR), New Delhi and Department of Agricultural Research and Education (DARE), Government of India.
Keywords: Corchorus, DNA extraction, microsatellite, mucilage, PCR, restriction digestion.
A simple miniprep based on early elimination of highly acidic and proteinaceous mucilages through ethanol washing of the tissue homogenates has been developed for the extraction of genomic DNA from mature leaves and seeds of Corchorus spp. As compared to high cetyltrimethylammonium bromide (CTAB)-NaCl DNA extraction followed by ethanol-based removal of remnant mucilages from the DNA pellet, this simple miniprep consistently and reproducibly recovers high amounts of DNA with good spectral qualities at A260/A280 and A260/A230. The purified DNA is efficiently digested by restriction endonucleases, and is suitable for PCR amplification of nuclear microsatellites with expected allele sizes.