PMC:4162895 / 5903-6899 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/4162895","sourcedb":"PMC","sourceid":"4162895","source_url":"https://www.ncbi.nlm.nih.gov/pmc/4162895","text":"Nineteen ISSR primers (Eurofins, Bangalore, India) were selected for the present investigation. PCR for amplifying the DNA preparations was carried out in a 25-μl volume of reaction mixture. A reaction tube contained 25 ng of DNA, 1 U of Taq DNA polymerase enzyme, 2.5 mM of each dNTPs, 1× Taq buffer with 25 mM MgCl2 and 25 pmol primers. Amplifications were carried out in a DNA thermal cycler (Eppendorf, mastercycler gradient) using following parameters: 94 °C for 5 min; 35 cycles at 94 °C for 1 min, 49–59 °C for 1 min, and 72 °C for 2 min; and a final extension at 72 °C for 10 min. The annealing temperature was adjusted to a range of 49–59 °C depending on GC content and length of the primers. PCR products were subjected to agarose gel [1.5 % (w/v)] electrophoresis in 1× TBE buffer, along with 1 kb DNA ladder (Fermentas Life Sciences) as size markers. DNA was stained with ethidium bromide and electrophoretic profile was photographed on gel documentation system (Alpha Innotech, USA).","tracks":[]}