CAJ | 학술논문

该文研究了蜘蛛大分子量基因组DNA(HMW-gDNA)的提取以及一种高效电洗脱纯化装置的构建.以蜘蛛胸部肌肉组织为原料,通过白改良CTAB法提取蜘蛛HMW-gDNA,利用透析膜和2 mL离心管构建一种新的HMW-gDNA快速凝胶回收装置,并对蜘蛛HMW-gDNA进行电洗脱分离回收.结果显示,改良CTAB法可高效提取蜘蛛HMW-gDNA(＞48.5 kb),且通过透析膜的截留作用,对普通琼脂糖凝胶中目的HMW-gDNA进行快速电洗脱分离,其回收率超过75%,OD_(260)/OD_(280)处于1.8～2.0之间,对HMW-gDNA完整性无影响.综合结果表明,改良CTAB法可用于蜘蛛HMW-gDNA的提取,此电洗脱纯化装置可从普通琼脂糖中高效回收HMW-gDNA,是一种低成本、简捷、高效且实用性强的凝胶回收方法.
해문연구료지주대분자량기인조DNA(HMW-gDNA)적제취이급일충고효전세탈순화장치적구건.이지주흉부기육조직위원료,통과백개량CTAB법제취지주HMW-gDNA,이용투석막화2 mL리심관구건일충신적HMW-gDNA쾌속응효회수장치,병대지주HMW-gDNA진행전세탈분리회수.결과현시,개량CTAB법가고효제취지주HMW-gDNA(＞48.5 kb),차통과투석막적절류작용,대보통경지당응효중목적HMW-gDNA진행쾌속전세탈분리,기회수솔초과75%,OD_(260)/OD_(280)처우1.8～2.0지간,대HMW-gDNA완정성무영향.종합결과표명,개량CTAB법가용우지주HMW-gDNA적제취,차전세탈순화장치가종보통경지당중고효회수HMW-gDNA,시일충저성본、간첩、고효차실용성강적응효회수방법.
Extraction of high molecular weight genomic DNA (HMW-gDNA) from spider organs and construction of an electroelution system for obtaining HMW-gDNA efficiently are described. We improved the CTAB method to isolate genomic DNA from spider pectoral muscle tissues, constructed a novel electroelution system with the use of 2-mL centrifuge tube and dialysis membrane, and utilized this self-constructed electroelution system to recover target HMW-gDNA from agarose gels. The HMW-gDNA extracted using the improved CTAB method could be over 48.5 kb in size. By trapping the HMW-gDNA with dialysis membrane, the electroelution system could recover target HMW-gDNA from agarose gels rapidly and without affecting the integrity of HMW-gDNA. The recovery rate was over 75% with theOD_(260)/OD_(280) ranging from 1.8 to 2.0. The results demonstrate that the improved CTAB method can be used to extractspider HMW-gDNA, and the electroelution system is a general method for recovering target HMW-gDNA from agarose gels with low cost, simplicity, high efficiency and good practicability.