Orthogonal optimization of SCoT-PCR system and primer screening in pumpkin
WANG Fengfeng
WANG Ping
SHI Lei
YANG Jing
YANG Qin
Abstract：A L2S(56) of orthogonal design was used to optimize SCoT-PCR (Scodon Targeted Polymorphism Amplification System) of pumpkin in 5 factors such as Taq DNA polymerase,Mg2+,DNA template,dNTPs and primer concentrations.The result showed that the optimized system was as follows:a total volume of 20 μL including 2.50 mmol · L-1 Mg2+,0.40 mmol·L-1 dNTPs,0.50 U Taq polymerase,1.00 μmol· L-1 primer and 30 ng DNA template.20 primers were selected from the 51 primer combinations test with clear band patterns and abundant polymorphisms.The most suitable annealing temperature of primers were selected.The SCoT-PCR reaction system for SCoT markers of pumpkin could be applied in pumpkin molecular genetics research.
Keywords：PumpkinSCoT-PCRSystem optimizationOrthogonal designPrimers screening
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 12-17 )