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以荔枝幼果和离区为材料,以N-十二烷基肌氨酸钠(LSS)为RNA提取液的主要成分,建立了一种简单快速高效适合于荔枝幼果和离区总RNA提取的新方法,简称LSS法。该法提取RNA的过程只需2h,且质量高,幼果和果柄RNA的28S/18S值分别为1.7和1.9,RNA完整性(RIN)值分别为9.7和10.0,分别为380和450μg.g-1,A260/A280值分别为1.80和1.89。将提取的RNA经DNA酶消化后反转录成cDNA进行PCR,扩增出预期长度的目的片段,满足进行后续基因表达的要求。LSS法操作简单快速,对微量组织样品是一种有效的RNA提取方法。
In this study, young and isolated Litchi chinense L. were taken as the main materials and N-lauryl sarcosine (LSS) was used as the main component of RNA extract to establish a simple, The new method, referred to as LSS method. The method of RNA extraction only 2h, and the quality of young fruit and stalk RNA 28S / 18S values were 1.7 and 1.9, RNA integrity (RIN) values were 9.7 and 10.0, respectively, and 450μg. g-1, A260 / A280 values were 1.80 and 1.89 respectively. The extracted RNA was DNase digested and reverse transcribed into cDNA for PCR amplification of the desired fragment of the desired length to meet the requirements for subsequent gene expression. The LSS method is simple and fast to perform and is an effective RNA extraction method for micro-tissue samples.