Optimization of SRAP-PCR amplification system for Populus by uniform design

A uniform design was applied to optimize the SRAP-PCR (sequence-related amplified polymorphism-polymerase chain reaction) system in Populus. Five factors in this SRAP-PCR, including MgCl 2, primers, dNTPs, TaqDNA polymerase and a DNA template, were tested using two uniform designs U16(45 ) and U12(35 ). A suitable SRAPPCR system for Populus was established. To 25 μL SRAP-PCR amplification reaction solution, 3.5 mmol/L MgCl 2, 0.20 mmol/L dNTPs, 0.44 μmol/L primer, 1.50 U TaqDNA polymerase and a 28 ng/L template were added. The annealing temperature had no clear effect on the amplification results. By using this optimal system, genomic DNAs from 10 species of Populus were tested and DNA bands were clear and showed high polymorphism. It suggests that the improved SRAP-PCR system can be used to study genetic relationships, systematic evolution and genetic diversity in Populus. 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