Establishment and Optimization of RAPD-PCR Reaction System in Eggplant

The genomic DNA of eggplant(Solanum melongena) was extracted by the modified CTAB method.The factors affecting RAPD results were studied through single factor and multi level tests.An optimal RAPD-PCR reaction system(20 μL) for eggplant was established,and it contained 25 mmol/L MgCl2 2.0 μL,10×PCR Buffer 2.0 μL,10 mmol/L dNTP 0.5 μL,5 U/μL Taq E 0.2 μL,0.1 μmol/L Primer 3 μL,10 ng/L template DNA 3 μL and water 9.3 μL.The appropriate amplification procedure was as follows: pre-denaturing for 5 min at 94 ℃,denaturing for 1 min at 94 ℃,annealing for 1 min at 37 ℃,elongating for 1.5 min at 72 ℃,45 cycles;elongating for 10 min at 72 ℃ and preserving at 4 ℃.