三种测序DNA模板制备方法的比较

Comparison of three methods for preparing DNA sequencing templates

背景:DNA模板质量对DNA序列测定起着至关重要的作用。目的:为基因组DNA或甲基化DNA测序寻找一种经济,简便的方法。方法:分别采用96管集合板及96孔板提取质粒,并且针对质粒设计一对包含目的片段的引物,扩增后纯化PCR产物,通过以上3种方法制备DNA测序模板进行测序。结果与结论:实验所采用的3种方法对于基因组DNA测序效果无差异(P>0.05)。对于甲基化DNA测序效果,96管集合板法优于其他2种方法(P<0.05)。说明3种方法均适用于基因组DNA的测序,而96管集合板法更适用于甲基化DNA的测序。

BACKGROUND：The quality of DNA templates plays a crucial role in DNA sequencing.OBJECTIVE：To provide an economical,simple method for genomic DNA sequencing and methylated DNA sequencing.METHODS：Plasmid extraction was performed using NucleoSpin multi-96 plus plasmid kit and edgebio kit SeqPrep？ 96 HP plasmid prep system,respectively.Primer pairs containing target fragments were designed for the plasmids.PCR products were purified after amplification.DNA sequencing templates were prepared by the three methods above.RESULTS AND CONCLUSION：There was no significant difference in the genomic DNA sequencing results between the three methods used in the experiment（P 0.05）.As for the methylated DNA sequencing,the NucleosPin multi-96 plus plasmid kit was better than the other two methods（P 0.05）.These findings indicate that the three methods are suitable for genomic DNA sequencing,while the NucleosPin multi-96 plus plasmid kit is more situable for the methylated DNA sequencing.