摘要
水体沉积物为一新型微生物种质资源库,不依赖于培养的菌种多样性研究的首要条件是获取优质的基因组DNA。本研究以改进的氯仿异戊醇法、蛋白酶K+SDS法、玻璃珠+蛋白酶K+SDS法提取湖泊基因组DNA,并与试剂盒提取结果进行比较,为沉积物分子生态学的研究提供参考和借鉴。结果表明,改进后的氯仿异戊醇法提取的DNA纯度较低,腐殖质明显,纯化后PCR产量较低;蛋白酶K+SDS法获得的DNA完整性较好,浓度高,PCR扩增应用效果最好;玻璃珠+蛋白酶K+SDS法提取的基因组DNA效果次之;试剂盒法提取的基因组DNA浓度较低,片段长度略小。蛋白酶K+SDS法为除试剂盒外适合于湖泊沉积物基因组DNA提取的较好方法。
Sediment is a novel and valuable microbial resources, and the extraction of excellent genomic DNA is the prerequisite for the culture-independent study of bacterial diversity. In this research, the improved chloroform/ isoamyl alcohol method, proteinase K＋SDS method and glass beads＋proteinase K＋SDS method were used to extract the genomic DNA from lake sediment samples. The extraction results were compared with that of the kit, providing references for the molecular ecology study of sediment. Results showed that the purity of DNA extracted by the improved chloroform/isoamyl alcohol extraction method was lower with obvious humus and lower PCR production; the integrity of the DNA extracted by the proteinase K＋SDS method was comparatively good with higher concentration and the best PCR amplification. The DNA extracted by the glass beads＋proteinase K＋SDS method came the second; the concentration of genomic DNA extracted by the kit method was lower, and fragment was slightly shorter. Besides the kit method, the proteinase K ＋SDS method was a relatively good method for extracting genomic DNA from lake sediment.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2017年第4期1551-1555,共5页
Genomics and Applied Biology
基金
国家自然科学基金项目(31370147
31400103)
河南省省属高校基本科研业务专项(2014YWQQ16)
河南省高校科技创新团队支持计划(15IRTSTHN019)共同资助
