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- Title
Optimization of ISSR-PCR System for Nerviliae fordii (Hance) Schltr. and Its Primer Screening.
- Authors
Yuqing ZHOU; Peilin DU; Peng FU; Feng XIE; Yuefeng WANG; Hua ZHU
- Abstract
[Objective] To establish and optimize the ISSR-PCR system satiable for Nerviliae fordii (Hance) Schltr. [Methods] Five factors affecting the ISSR-PCR system reaction of N. fordii were optimized by orthogonal test, which were dNTP, template DNA, primer. Mg2+ and Taq DNA polymerase. Single factors in the lest were analyzed by new multiple range method. [Results] The optimal reaction system for N. fordii was as follows: The 20 μL total reaction volume included 2 μL 10 × PCR buffer, 225 μmol/L dNTP, 60 ng template DNA, 0.4 μmol/L primer, 2.5 miml/L Mg2+, 1.0 U Taq DNA polymerase, and 20 μL total reaction volume. A total of 16 ISSR primers were screened from 100 ISSR primers, which was stable and had almndanl polymorphism. [Conclusions] The established ISSR-PCR reaction system for N. fordii was stable and reliable, and could be used for the genetic differentiation analysis of N. fordii.
- Subjects
MEDICINAL plants; GENETIC research; PLANT genetics; GENETIC polymorphisms in plants; DNA polymerases; MAGNESIUM ions
- Publication
Medicinal Plant, 2015, Vol 6, Issue 9/10, p33
- ISSN
2152-3924
- Publication type
Article