CAJ | 학술논문

以荸荠叶状茎为试验材料,采用改良的SDS法提取其基因组DNA,并对其RAPD反应体系进行优化,建立了荸荠的BAPD-PCR优化反应体系和程序.结果表明,提取的基因组DNA纯度和完整性较好,OD260/OD280值在1.8～2.0之间,DNA无降解现象,完全可以满足RAPD-PCR扩增要求.建立了荸荠RAPD反应体系:总体积为25μl,各有关成分的最佳浓度分别为25mmol/L Mg2+,1.OU Taq DNA聚合酶,0.2mmol/L dNTPs,1μmol/μl引物,1.5ng/μl DNA模板.PCR反应程序为:94℃预变性3rnin;94℃变性1min,37℃退火30s.72℃延伸60s,40个循环;最后72℃延伸10min.
이발제협상경위시험재료,채용개량적SDS법제취기기인조DNA,병대기RAPD반응체계진행우화,건립료발제적BAPD-PCR우화반응체계화정서.결과표명,제취적기인조DNA순도화완정성교호,OD260/OD280치재1.8～2.0지간,DNA무강해현상,완전가이만족RAPD-PCR확증요구.건립료발제RAPD반응체계:총체적위25μl,각유관성분적최가농도분별위25mmol/L Mg2+,1.OU Taq DNA취합매,0.2mmol/L dNTPs,1μmol/μl인물,1.5ng/μl DNA모판.PCR반응정서위:94℃예변성3rnin;94℃변성1min,37℃퇴화30s.72℃연신60s,40개순배;최후72℃연신10min.
The young phylloclade of water chestnut was used as experimental material, and the improved SDS method was used to extract the genomic DNA of young phylloclade successfully. The results showed that the extracted genomic DNA was pure and integral, and the ratio of OD260/OD280 was from 1.8 to 2.0, and the degradation of extracted genomic DNA was not found. Therefore, the extracted genomic DNA was suitable for RAPD-PCR reaction. The conditions for RAPD-PCR were optimized, and the best RAPD reaction system was established as follows: total volume being 25μl, containing 25 mmol/L Mg2+, 1.0 U Tat/DNA polymerase, 0.2 mmol/L dNTPs, 1 μmol/μl primer and 1.5 ng/μl template DNA. The PCR program was 3 rain at 94℃ for predenaturalization, then 40 cycles of 1 min at 94℃ for denaturation, 30s at 37℃ for annealing and 60s at 72℃ for extension, finally extension at 72℃ for 10min