CAJ | 학술논문

快速而稳定地提取大草履虫的基因组DNA是进行后续分子生物学研究的前提。文章通过比较传统SDS法、CTABⅠ法和CTABⅡ法等提取大草履虫DNA的效果，进一步优化提取方法，获得快速稳定地提取大草履虫基因组DNA的方法。结果表明：用CTAB I法提取的DNA浓度低，杂质较多；CTABII提取的DNA电泳时检测不出条带；而SDS法提取的DNA在电泳时带型较整齐，无降解，且DNA的得率和纯度较高，但仍有蛋白质污染。针对蛋白质的去除，用SDS-蛋白酶K法和SDS-NaAc法来提取DNA，经过试验发现这2种方法都能够很好地去除蛋白质，所获DNA样品的纯度和得率都能用于进一步分子试验，而SDS-NaAc法更经济适用。
쾌속이은정지제취대초리충적기인조DNA시진행후속분자생물학연구적전제。문장통과비교전통SDS법、CTABⅠ법화CTABⅡ법등제취대초리충DNA적효과，진일보우화제취방법，획득쾌속은정지제취대초리충기인조DNA적방법。결과표명：용CTAB I법제취적DNA농도저，잡질교다；CTABII제취적DNA전영시검측불출조대；이SDS법제취적DNA재전영시대형교정제，무강해，차DNA적득솔화순도교고，단잉유단백질오염。침대단백질적거제，용SDS-단백매K법화SDS-NaAc법래제취DNA，경과시험발현저2충방법도능구흔호지거제단백질，소획DNA양품적순도화득솔도능용우진일보분자시험，이SDS-NaAc법경경제괄용。
Isolation of genomic DNA is the precondition for molecular research on Paramecium caudatum. Traditional SDS method,CTABⅠmethod and CTABⅡmethod were employed to extract the genomic DNA from P. caudatum,Based on the experimental results,an optimized extraction procedure was established. The results showed that the DNA extracted by CTAB I method was poor in both purity and yield. The DNA extracted by CTABⅡ method was even undetectable,and the DNA extracted by the traditional SDS method performed better in yield and purity,but there were also protein impurities in DNA samples. In order to re-move protein impurities,proteinase K and NaAc were separately added to treat the DNA samples extracted by the traditional SDS method. These two reagents could effectively remove protein impurities,and the yield-ed DNA could satisfy the requirements of molecular research in both purity and yield. Considering the experi-mental cost,the SDS-NaAc method was more economical and can be used to extract the genomic DNA from P. caudatum.