RT Journal
T1 OPTIMIZATION OF DNA EXTRACTION AND PCR PROTOCOL FOR RAPD ANALYSIS OF TOR PUTITORA
A1 N. Shafi
A1 S. Janjua
A1 S. Jafar K. Anwar
A1 A. Mian
JF Journal of Animal and Plant Sciences
JO JAPS
SN 1018-7081
VO 25
IS 3
SP 270
OP 275
YR 2015
FD 2015/06/01
DO DOI NA
AB Genetic analysis of biological organisms is dependent on a high yield of genomic DNA samples. This study was designed for the optimization of DNA extraction and PCR protocol for RAPD analysis of golden mahseer (Pisces: Tor putitora) to find out the genetic diversity among its different populations. The extraction is a modified phenol-chloroform method involving addition of 40 µL of proteinase K during overnight lysis of samples and successive long-term chloroform- isoamylalcohol extractions. The yielded DNA showed clear thick bands on agarose gel and its quality was not affected by the period of preservation. The isolated DNA was used for randomly amplified polymorphic DNA (RAPD) analysis. The protocol was optimized based on higher concentration of MgCl (2.5 mM), lower concentrations of primers (0.25 ρmol) 2 and Taq polymerase (0.02 units), 50 ng of template DNA and an annealing temperature of 34°C. Thus on the basis of results this protocol for DNA isolation and PCR was accurate for the genetic diversity analysis of different populations of Masheer. sequencing or any other form of the molecular
K1 Mahseer, Tor putitora, DNA extraction, PCR optimization, reaction parameters
PB Pakistan Agricultural Scientists Forum
LK https://thejaps.org.pk/AbstractView.aspx?mid=2015-JAPS-643