Optimization of SRAP-PCR System and Primers Screening for Rambutan (Nephelium lappaceum L.)
To establish a stability and reliable SRAP system for rambutan (Nephelium lappaceum L.),in this study,the PCR procedure was firstly evaluated by using a single-factor experiment,which involved various concentration levels of DNA template,Mg2+,dNTPs,Taq DNA polymerase and primers.Based on the initial results,another assay with orthogonal design of L16(45) was carried out,and eventually brought about an optimized system for rambutan as follows:a volume of 20 μL reaction mixture containing 20 ng genome DNA,0.25 mmol/L dNTP mixture,0.60 μmol/L each primer,1.0 U rTaq DNA polymerase and 2.5 mmol/L Mg2+.And then,37 primer pairs were screened out from 116 SRAP combinations based on their stable amplification,clear banding patterns and good polymorphism.In all,both the optimized SRAP-PCR reaction system and selected primers will facilitate the researches of genetic diversity and mapping in rambntan.
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NSTL
万方数据
维普
