Journal of Pure and Applied MicrobiologyVol. 9 No. 3

Use of CTAB Method for Isolation of Good Quality and Quantity of DNA

D. Ashwini1 and S.P. Tiwari2

1Department of Plant Pathology, JNKVV, Jabalpur, India. 2 Principal Scientist, Department of Plant Pathology, JNKVV, Jabalpur, India.

Received on 09 April 2015 and accepted on 13 June 2015

 

ABSTRACT

Isolation of good quality and quantity of DNA is essential for the purpose as well as for preserving the same to a considerable period of time. Although the DNA needed per reaction in PCR based makers is very low, the number of PCR reactions to be performed is large and hence agood quality and quantity of DNA would be needed for such studies. We have CTAB (Cetyl tri methyl ammonium bromide) method for isolation of DNA from plant tissue. The method is suitable for isolating DNA from a small to medium number of plant samples. The DNA can be stored for a longer duration. The method involves extraction of DNA using a buffer (pH 8.0) containing Tris HCL (100 mM), EDTA (20mM), 5 M NaCl, 10% CTAB and 2% b- mercaptoethanol, followed by purification of DNA with phenol, chloroform and Isoamly alcohol and finally precipitation of DNA by sodium acetate and isopropanol. The protocol is simple and does not require expensive chemicals such as proteinase K, liquid nitrogen etc.

Keywords : Isolation of DNA, CTAB method.