African Journal of Biotechnology Vol. 5 (8), pp. 585-589, 18 April 2006   

ISSN 1684–5315 © 2006 Academic Journals        

Full Length Research Paper

Small and large scale genomic DNA isolation protocol for chickpea (Cicer arietinum L.), suitable for molecular marker and transgenic analyses

Dipankar Chakraborti, Anindya Sarkar, Sumanti Gupta and Sampa Das^*.

Plant Molecular and Cellular Genetics, Bose Institute, P1/12 C.I.T. Scheme VIIM, Kolkata –7000054, India.

. *Corresponding authors E-mail. sampa@bic.boseinst.ernet.in.

Abbreviations: CTAB, Hexadecyltrimethylammonium bromide; ISSR, inter-simple sequence repeat; PCR, polymerase chain reaction; Tris, tris (hydroxymethyl)-aminomethane.

Accepted 27 January, 2006.

Chickpea is an important food legume crop with high nutritional value. Lack of appropriate DNA isolation protocol is a limiting factor for any molecular studies of this crop. The present report describes a rapid and efficient protocol for small and large scale preparation of superior quality and quantity of DNA from four cultivars (JG62, WR315, C235 and ICCV89314) compared to that of earlier reports. The yield of DNA through both the methods was estimated to be approximately 80 μg per g of plant tissue. Both small and large scale preparations were essentially suitable for PCR and Southern blot hybridization analyses, which are the key steps in crop improvement programme through marker development and genetic engineering techniques.

Key words: Cicer arietinum L., phenolics, restriction enzyme digestion, PCR amplification, Southern hybridization.