The optimal ISSR-PCR reaction conditions in tea was established by studying the main parameters. Results showed that the optimum concentration of five important components i.e. template DNA, primer, Mg++, dNTP, Taq DNA polymerase in 20βµl reaction mixture was 10ng, 150βnmol/L, 1.5βmmol/L, 150βµmol/L, 0.5βU, respectively. The appropriate annealing temperature was average 4.5℃ higher than Tm of corresponding primer, and at least 30 PCR cycles should be carried out to ensure sufficient PCR products. ISSR polymorphism between thirteen tea germplasm was 77.6%, and 12 out of 13 tea germplasm could be identified by ISSR fingerprinter established with primer TRI18.
YAO Ming-zhe
,
WANG Xin-chao
,
CHEN Liang
,
YANG Ya-jun
. Establishment of ISSR-PCR Reaction Conditions in Tea Plant[J]. Journal of Tea Science, 2004
, 24(3)
: 172
-176
.
DOI: 10.13305/j.cnki.jts.2004.03.005
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