Primers for
Ustilagospp. 5.8S-18S-28S rRNA Intergenic spacer region
were used for screening (Forward Primer(S1) 3-GCAGC
CGATAATCTACCAA-5 and Reverse Primer- (S2)
5-CCAGCTTCTTGCTCATCCTC-3). The PCR conditioninitial denaturation at 94C for 5 min, followed by 35
cycles as follows: Denaturation-94C for 30 s, Primer
annealing-56C for 30 s, Primer Extension-72C for
60 s, Final Extension 72C for 5 min, The PCR reaction
was carried out with 2ll plant DNA and total volume of
reaction mixture was 25ll. The amplicons were analyzed
on 0.8% agarose gel as described by Sambrook et al.
(1989) and the bands visualized under UV light after
staining with ethidium bromide