Melon(Cucumis Melol) Varieties Fingerprint Construction And Molecular Markers Application In Hybrid Purity Detection

Since twenty-first Century has been the rapid development of molecular biology, in basic research more deeply at the same time in its application is more and more widely, molecular marker technology is one of them. In this paper the application of restriction site amplified polymorphism (Restriction Site Amplification Polymorphism, RSAP) labeling technique to the study of Melon Hybrid seed purity identification, and discussed RSAP technology optimization, and success of sweet melon Lou No.1of purity identification.1) using L16(45) orthogonal experiment, the PCR system relates to5factors set4level gradient in all16combination, repeated two times each combination set. The final optimized melon PCR amplification system was25μL:10×buffer2.5μL,0.25mmol/L primers for dNTPs,0.25μmol/L,1.5mmol/LMg C12, DNA template6.5ng/μ L, Taq enzyme2.0U.2) optimized RSAP reaction procedures:94℃for5minutes;94℃50s,35℃50s72℃100s5cycle;94℃50s,45℃50s,72℃100s35cycles; the last extension5min.3) established on Lou Tianyi melon fingerprint, wherein the primer R2R5can successfully be Lou Tianyi number hybrid hybrid detected, its results and field testing results coincide with the rate of93%.