Abstract: The orthogonal design table of L16(45)was used to optimize the five factors(Taq DNA polymerase,Mg2+,DNA template,dNTPs and primer concentrations)of SCoT-PCR(start codon targeted polymorphism)system in tree peony.The results showed that the optimized system was as follows:a total volume of 20 μL including 1.0 ng/μL DNA template,0.50 U Taq polymerase,0.60 μmol/L primer,2.50 mmol/L Mg2+,0.25 mmol/L dNTPs and 1 PCR Buffer.Each factor had different effect on the results.The concentration of Mg2+ was the key factor affecting the SCoT-PCR system.The optimized SCoT-PCR system was tested on seventeen peony genus,and the result was stable and reliable.From the 36 primer combinations tested,Twenty-four were selected with clear band patterns and abundant polymorphism.The optimized SCoT-PCR system and polymorphism primmer combinations provided the basis for further research on Paeonia suffruticosa.

Key words: SCoT-PCR, Reaction system, Orthognal design, Primers screening

CLC Number: 

• S685.11