Selection of primers and establishment of SSR-PCR reaction system on Pinus tabulaeformis Carr

Taking Pinus tabulaeformis Carr.as the material,the effects of primer pairs,whole DNA concentration,Mg²⁺and Taq polymerase concentration and dNTP concentration on SSR reaction system were studied.Twelve primer pairs with abundant polymorphism bands were selected and the SSR-PCR reaction system on P.tabulaeformis was established too.The results showed that in 15 μL PCR reaction:30 ng DNA,(0.25 mmol/L) Mg²⁺,0.2 mmol/L dNTP,250 nmol/L Primer(F),250 nmol/L Primer(R),0.375 U Taq polymerase were the best.Detected by 6% polyacrylamide gel,polymorphism among 49 clones of P.tabulaeformis in the Seed Orchard of Xingcheng,Liaoning Province was abundant, the length of amplification products was 100-250 bp,the number of alleles per locus varied from five to ten.The work provides a powerful tool for paternity analysis in P.tabulaeformis seed orchards.