Study on optimization of SRAP-PCR reaction system for Pinellia ternata in Suzhou.
- Author:
Ai-Min ZHANG
1
;
He-Dong LU
;
Jian-Ping XUE
;
Xing-Kui TAO
;
Tao XUE
;
Wei SHENG
;
Yan-Fang ZHU
Author Information
- Publication Type:Journal Article
- MeSH: China; DNA Primers; genetics; DNA, Plant; genetics; Electrophoresis; Magnesium; metabolism; Nucleic Acid Amplification Techniques; methods; Nucleotides; genetics; Pinellia; genetics; Polymerase Chain Reaction; methods; Reproducibility of Results; Taq Polymerase; metabolism; Templates, Genetic
- From: China Journal of Chinese Materia Medica 2012;37(24):3815-3818
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the optimization system of SRAP-PCR molecular marker technology in the analysis on Pinellia ternata.
METHODSRAP-PCR reaction system for P. ternata was optimized by L16 (5(4)) orthogonal design with five elements (dNTPs, Mg2+, the template DNA, primers, Taq enzyme) and four standards.
RESULTThe most suitable forward primer for SRAP for Pinellia ternata was 5'-TGAGTCCAAACCGGAAG-3', while the reverse primer was 5'-GACTGCGTACGAATTACG-3'. The optimized reaction system contained 70 ng DNA template, 0.9 micromol x L(-1) primer, 0.20 mmol x L(-1) dNTP s, 1.5 - 2.0 mmol x L(-1) Mg2+, and 2 U Taq enzyme.
CONCLUSIONSRAP-PCR system for P. ternata is established to lay a foundation for future construction of SRAP genetic map of P. ternata.
