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WANG Chen-Ying, ZHAO Jian-Cheng. Optimization of ISSR-PCR Reaction System in a Rare Medicinal Plant Rhodobryum ontariense(Bryaceae)[J]. Plant Science Journal, 2009, 27(6): 679-683.
Citation: WANG Chen-Ying, ZHAO Jian-Cheng. Optimization of ISSR-PCR Reaction System in a Rare Medicinal Plant Rhodobryum ontariense(Bryaceae)[J]. Plant Science Journal, 2009, 27(6): 679-683.

Optimization of ISSR-PCR Reaction System in a Rare Medicinal Plant Rhodobryum ontariense(Bryaceae)

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  • Received Date: December 02, 2008
  • Revised Date: April 06, 2009
  • Published Date: December 25, 2009
  • To seek a standardizing program of ISSR technique for the genetic diversity analysis of a rare medicinal plant Rhodobryum ontariense,a single-factor experiment was designed to optimize ISSR-PCR amplification system.A suitable ISSR reaction system was obtained,that is:20 μL reaction system containing 6 ng DNA template,0.4 μmol/L primer,2.25 mmol/L Mg2+,0.6 U Taq DNA polymerase,0.4 mmol/L dNTPs.The profile of ISSR amplification was 4 min at 94℃,followed by 40 cycles of denaturation for 1 min at 94℃,annealing for 1 min at 48~50℃(according to specific primer),and extension for 1 min at 72℃,and ending with a final extension for 7 min at 72℃,stored at 4℃.
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