Optimizing of the PCR Amplification Conditions and Sequencing the Weedy Rice nrDNA ITS

doi:10.7668/hbnxb.2008.06.039

Abstract

Abstract: The PCR conditions of the weedy rice nrDNA ITS were optimized in this study,as a result,the PCR reactions were carried out as follows:each reaction mixture contained 1μL genomic DNA,0.125 mmol/L each dNTPs,0.5mmol/L forward and reverse primer,2 10μL 10@PCR reaction Buffer,and the f inal react ion was adjusted to 20 μL with sterilized ddH 2O.The annealing temperature was 57℃.It can assure that PCR products quality and purity using these optimal conditions.The length of the ITS fragments was about 600 bp using the direct sequencing methods and consistent with the genebank data.The phylogenetic information of the weedy rice ITS would conveniently provide the direct molecular evidence to clarify its origin and evolution.

Key words: Weedy rice, nrDNA ITS, PCR amplification conditions, Sequencing

CLC Number:

S511.01

LIU Zhi-wen, HAN Xu, ZHOU Suo-tong, WANG Li, LI Xian-zhen, CHEN Wen-fu. Optimizing of the PCR Amplification Conditions and Sequencing the Weedy Rice nrDNA ITS[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(6): 168-170. doi: 10.7668/hbnxb.2008.06.039.

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