Authors
Parthadeb Ghosh, Sanjib Kumar Chattopadhyay, Sinchan Adhikari, Soumen Saha, Subhasis Mondal, Abdul Kader, Soura Tapas Dey, Tapas Kumar Bandyopadhyay, and Chandan Sengupta
Summary
This method is designed to recover utmost amount of DNA from heterogeneous plant tissues restraining the co-extraction of PCR inhibiting substances. It requires maceration of plant tissue of about 1.0 cm2 in DNA extraction buffer [200 mM Tris-HCl (pH 8.0), 200 mM NaCl and 25 mM EDTA, and 1% PVP], followed by cell lysis with 10% SDS. The lysate was treated with Phenol: chloroform: isoamyl alcohol (25:24:1) efficiently removes all chaotropic cellular materials except water soluble polysaccharides which concomitantly excluded by DNA precipitation with ethanol and sodium chloride. It needs about one and half an hour to prepare DNA to perform thousands of PCR-based reactions and other DNA manipulation techniques. This method does not require liquid nitrogen and also bypass RNase treatment. It can be performed even in low technology laboratories.Further details
The protocol was published on Protocol Exchange on 7 February 2013. To see the entire protocol, click on the source link.Advertisement
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