Establishment of TRAP-PCR Reaction System and Development Strategy of Primers for Beet

doi:10.11924/j.issn.1000-6850.casb15080024

Abstract

Abstract: With sugar beet as the material, the influence of concentration changes of the four factors of dNTPs, primer, template and DNA polymerase in the beet TRAP-PCR reaction system on the amplification results was discussed with single-factor experiment, and finally the optimal TRAP-PCR reaction system of sugar beet was established. The results showed that in the 20 μL reaction system, the beet TRAP-PCR optimal reaction system contained 2.0 μL of 10×PCR buffer (including Mg²), 10 ng of template DNA, 0.75 U of Taqase, respectively 0.8 μL of 10 μmol/L fixed primer and random primer, and 0.5 μL of dNTPs (2.5 mmol/L each). In addition, a method for rapid development of beet TRAP-PCR primer was proposed, 15 varieties of sugar beer were amplified with the optimized program. The results show that the system has stable amplification results, clear bans and abundant polymorphism of partial primer, and can be used for the construction of sugar beet variety fingerprint and research in other fields of molecular biology.

CLC Number:

S566.3

Wu Zedong,Liu Naixin,Qin Haodong,Ni Hongtao and Ma Longbiao. Establishment of TRAP-PCR Reaction System and Development Strategy of Primers for Beet[J]. Chinese Agricultural Science Bulletin, 2016, 32(6): 96-101.

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