CAJ | 학술논문

以短序大功劳嫩叶为材料，采用CTAB法、CTAB改良法1、CTAB改良法2、SDS法和试剂盒法五种方法提取短序十大功劳基因组总DNA，用分光光度计和琼脂糖凝胶电泳方法检测所得总DNA的纯度和得率，用ISSR-PCR扩增的方法检测所得总DNA的质量。结果表明，五种方法均能从短序大功劳叶片中提取到基因组DNA，但不同方法提取得的基因组DNA的纯度、浓度和得率存在明显的差异。CTAB改良法2和试剂盒法提取的DNA纯度高，可直接用于下游分子生物学实验，CTAB法、CTAB改良法1和SDS法提取的总DNA质量较差，不利于下游的分子生物学实验；五种方法提取的总DNA的得率在10.836~451.709μg/g之间，呈CTAB法>SDS法>CTAB改良法1>CTAB改良法2>试剂盒法的现象。此实验获得的结果可以为短序十大功劳分子生物学研究提供基础。
이단서대공로눈협위재료，채용CTAB법、CTAB개량법1、CTAB개량법2、SDS법화시제합법오충방법제취단서십대공로기인조총DNA，용분광광도계화경지당응효전영방법검측소득총DNA적순도화득솔，용ISSR-PCR확증적방법검측소득총DNA적질량。결과표명，오충방법균능종단서대공로협편중제취도기인조DNA，단불동방법제취득적기인조DNA적순도、농도화득솔존재명현적차이。CTAB개량법2화시제합법제취적DNA순도고，가직접용우하유분자생물학실험，CTAB법、CTAB개량법1화SDS법제취적총DNA질량교차，불리우하유적분자생물학실험；오충방법제취적총DNA적득솔재10.836~451.709μg/g지간，정CTAB법>SDS법>CTAB개량법1>CTAB개량법2>시제합법적현상。차실험획득적결과가이위단서십대공로분자생물학연구제공기출。
In this paper, five different methods of DNA extraction, i.e. CTAB method, improved CTAB method 1, improved CTAB method 2, SDS method and Kit method, were used to isolate genomic DNA from the fresh tender leaves materials of Mahonia breviracema Y.S. Wang et Hsiao. And then, the spectrophotometer, agarose gel elec-trophoresis and ISSR-PCR amplification were respectively used to investigate the genomic DNA extraction effects. The results showed that, all the five methods could extract the genomic DNA from the leaves of M. breviracema, but there were obvious differences in extracted purity and yield of genomic DNA by with different methods. The CTAB method 2 and Kit method could yield relatively pure total DNA which were highly suited for use directly in down-stream applications. Whereas the CTAB method, improved CTAB method 1and SDS method yield genomic DNA of poor quality which could not be used in subsequent PCR application directly. The genomic DNA yielded with five methods was ranged from 10.836μg/g to 451.709μg/g and was in order as the CTAB method>SDS method>CTAB method 1>CTABCTAB method 2>Kit method. The results could provide some basis for the molecular research of M. breviracema.