Short communication PCR amplification of the genomic DNA from the seeds of Ceylon ironwood, Jatropha, and Pongamia Vigya Kesari, Medhavi Sudarshan, Archana Das, Latha Rangan* Department of Biotechnology, Indian Institute of Technology Guwahati, North Guwahati 781 039, Assam, India article info Article history: Received 1 October 2008 Received in revised form 31 March 2009 Accepted 9 August 2009 Available online 27 August 2009 Keywords: DNA isolation Jatropha curcas Mesua ferrea Pongamia pinnata RAPD PCR Restriction enzymes Taq DNA polymerase abstract Plant source for fuel that replaces fossil fuels is a topical subject and has gained prominence as ‘‘Biofuel crops’’. Successful DNA extraction from seed yielding appropriate DNA quality for PCR amplification will allow molecular genetic investigations in such crops. Standardized protocols for DNA isolation failed to yield high quality DNA from dried seeds that are rich source of triglycerides. In this paper, we report a protocol for isolation of genomic DNA from three potential biofuel crops using SDS extraction step, followed by precipitation and purification to remove polysaccharides, proteins and polyphenols which are abundant in storage tissues like seeds. The average yield of DNA among the biofuel crops varied from 15 to 25 mg kg 1 tissue. Spectrophotometric and electrophoretic analysis indicated that the isolated DNA was highly pure and of high molecular weight amenable for PCR amplification and restriction endonucleases. This procedure may prove useful for other oilseed crops of commercial importance. ª 2009 Elsevier Ltd. All rights reserved. 1. Introduction DNA marker technology has been rapidly developing and many techniques that seemed unfeasible before are now routinely used. In anticipation of new marker technology, we have been exploring basic approaches for extracting whole genomic DNA from dried seeds, as a part of a case study on a biodiesel plants (Pongamia pinnata, Mesua ferrea and Jatropha curcas). The ability to identify relevant gene(s) involved in fatty acid biosynthesis and the ability to reliably extract good quality DNA from seed samples is a fundamental step in the application of genetic techniques to the success of biofuel crops. So far, the seeds which are rich source of triglycerides had very little attention from plant molecular biologists. The isola- tion of high quality DNA is a prerequisite for any molecular biological studies when using the seed source. The seeds of these plants contain exceptionally high amount of poly- saccharides, polyphenols, and other secondary metabolites that can hamper DNA isolation, amplification, restriction digestion and subsequent molecular cloning [1–5]. Polysaccharides and phenolic compounds bind to nucleic acids during DNA isolation and interfere with subsequent reactions [6]. A number of methods are available and are being devel- oped for the isolation of nucleic acid from seeds and Abbreviations: EDTA, ethylenediaminetetraacetic acid; EtBr, Ethidium bromide; PCR, Polymerase chain reaction; RAPD, Random amplification of polymorphic DNA; SDS, Sodium dodecyl sulphate; TE, tris–EDTA buffer. * Corresponding author. Department of Biotechnology, Indian Institute of Technology Guwahati, Room No. 202, O Block, Guwahati 781 039, Assam, India. Tel.: þ91 361 2582214; fax: þ91 361 2582249. E-mail address: latha_rangan@yahoo.com (L. Rangan). Available at www.sciencedirect.com http://www.elsevier.com/locate/biombioe 0961-9534/$ – see front matter ª 2009 Elsevier Ltd. All rights reserved. doi:10.1016/j.biombioe.2009.08.005 biomass and bioenergy 33 (2009) 1724–1728