TY - JOUR
T1 - Optimizing System of SSR-PCR in Pinus radiata and Pinus tabulaeformis
AU - Zong-Xing, Wu AU - Jing-Yong, Zhong AU - Fan, Zhang AU - Cheng-De, Luo AU - Lan, LuXiu- AU - Mei, LI
JO - Journal of Molecular Genetics
VL - 1
IS - 2
SP - 44
EP - 49
PY - 2009
DA - 2001/08/19
SN - 2070-4267
DO - jmolgene.2009.44.49
UR - https://makhillpublications.co/view-article.php?doi=jmolgene.2009.44.49
KW - optimization
KW -SSR
KW -Pinus tabulaeformis
KW -Pinus radiate
KW -reaction system
KW -China
AB - Reaction system of SSR-PCR was optimized by orthogonal design using 21 Pinus radiate and 3 Pinus tabulaeformis originated from the different locations. The results indicated that the optimal reaction system of SSR for P. radiata and P. tabulaeformis were described as follows each DNA amplification was carried out in a volume of 25 μL containing 0.3 U Taq polymerase, 1.5 μL Mgcl2 (2.0 mmol L-1), 160-200 ng genomic DNA, 0.3 μL each of SSR forward and reverse primers (0.6 μmol L-1) and 2.0 μL dNTPs (0.2 mmol L-1). DNA amplification for SSR was performed for initial 3 min at 94°C for pre-denaturing, followed by 30 cycles at 94°C for 30 sec for denaturing, 45 sec at 45-49°C for annealing, 30 sec at 72°C for extension. The reaction was terminated with 5 min extension at 72°C. The reaction system was verified by using 24 materials. The numbers of alleles per primer were 5-10 and the range of polymorphic bands was 100-250 bp.The results suggested that the reaction system described as above exhibited high polymorphic percentage and good stability and repetition.
ER -