CAJ | 학술논문

用修改了的异硫氰酸胍法从两种森林昆虫Clostera anastomosis 和 Saperda populnea 的不同组织中快速提取总RNA.电泳检测和cDNA合成分析结果表明,RNA未降解.经紫外分光光度计检测,A260/ A280在1.8 到 2.0之间,表明RNA的纯度很高.用RT-PCR合成的双链cDNA的长度大于2kb,表明mRNA的完整性.用PCR的方法克隆了C. anastomosis β肌动蛋白和几丁质酶的基因片段,表明RNA可用于其它分子操作.使用这个方法,在4个小时内可至少从8个样品中提取RNA并进行电泳分析.这些结果表明用这个修改后的一步法提取昆虫总RNA是省时,省钱和有效的.
용수개료적이류청산고법종량충삼림곤충Clostera anastomosis 화 Saperda populnea 적불동조직중쾌속제취총RNA.전영검측화cDNA합성분석결과표명,RNA미강해.경자외분광광도계검측,A260/ A280재1.8 도 2.0지간,표명RNA적순도흔고.용RT-PCR합성적쌍련cDNA적장도대우2kb,표명mRNA적완정성.용PCR적방법극륭료C. anastomosis β기동단백화궤정질매적기인편단,표명RNA가용우기타분자조작.사용저개방법,재4개소시내가지소종8개양품중제취RNA병진행전영분석.저사결과표명용저개수개후적일보법제취곤충총RNA시성시,성전화유효적.
A modified guanidinium isothiocyanate method was used to extract total RNA from two forest insect species Clostera anastomosis and Saperda populnea. The integrity of RNA was demonstrated by the methods of gel electrophoresis and cDNA analysis. Typical A260/ A280 absorbance ratio of the total RNA was in range of 1.8 to 2.0. The size of double strand cDNAs obtained by RT-PCR was more than 2 kb, which indicated that intact mRNA was obtained. The fragments of β-actin and chitinase gene from the RNA of C. anastomosis were obtained by RT-PCR, which indicated that the RNA could be used for other molecular operation. By this procedure, RNAs could be extracted and analyzed by electrophoresis from at least 8 samples within 4 hours. These results showed that this method was time- and cost-saving and effective.