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首页> 外文会议>International Conference on Biological Science >Optimization of the CTAB3-LiCl and commercial kit methods in the process of RNA isolation and amplification in strawberries fruit (Fragaria spp.)
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Optimization of the CTAB3-LiCl and commercial kit methods in the process of RNA isolation and amplification in strawberries fruit (Fragaria spp.)

机译：CTAB3-LICL和商业试剂盒方法在草莓果实RNA分离和扩增过程中的优化（Fragaria SPP。）

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RNA (Ribonucleic Acid) isolation is a basic technique in the field of molecular biology. The purpose of RNA isolation is to acquire pure mRNA that can be used to see the expression of certain genes in an individual. Many methods can be used to perform RNA isolation, both conventional and modern methods. The purpose of this study were to compare the results of total RNA isolated using the CTAB_3-LiCl (conventional) method and the Commercial Kit (modern) method and then amlification of FaPYR1 and FaCHS genes from mRNA templates in the strawberry fruits. The RNA isolation done by using the Kit method referring to the Geneaid "RNA Total Mini Kit (Plant)" kit protocol. In general, the RNA isolation may consist of three stages: (1) Lysing the membrane or cell wall by physical and chemical destruction, (2) Removing unwanted components such as proteins, polysaccharides, and cell debris, (3) Searching for pure RNA by precipitation. The total RNA isolates were analyzed quantitatively by TECAN nanodrop spectrophotometer to determine the purity and concentration of RNA. Biorad thermal cylers used for amplification for the target genes. The results showed that the RNA isolated using the CTAB_3-LiCl method had a higher RNA concentration than the Kit method, and the purity produced from the two methods did not differ significantly. Thus, in this research, the total RNA isolated using the CTAB_3-LiCl method is more optimal than the Kit method. The FaPYR1 and FaCHS genes amplificated in 627 bp, Actin gene in 262 bp, and 26S-18S Housekeeping gene in 146 bp to all samples selected.
机译：RNA（核糖核酸）分离是分子生物学领域的基本技术。 RNA分离的目的是获得纯mRNA，可用于在个体中看到某些基因的表达。许多方法可用于进行常规和现代方法的RNA隔离。本研究的目的是使用CTAB_3-LICL（常规）方法和商业套件（现代）方法和商业试剂盒（现代）方法进行比较分离的总RNA的结果，然后从草莓果实中的mRNA模板融合FAPYR1和FACHS基因。通过使用试剂盒方法来进行RNA分离，所述试剂盒方法是指Gateaid“RNA总迷你试剂盒（植物）”试剂盒方案。通常，RNA分离可以由三个阶段组成：（1）通过物理破坏裂解膜或细胞壁，（2）除去不需要的组分，例如蛋白质，多糖和细胞碎片，（3）寻找纯RNA通过降水。通过Tecan Nanodrop分光光度计定量分析总RNA分离物，以确定RNA的纯度和浓度。 Biorad热缸用于靶基因的扩增。结果表明，使用CTAB_3-LICL方法分离的RNA具有比试剂盒方法更高的RNA浓度，并且由两种方法产生的纯度没有显着差异。因此，在本研究中，使用CTAB_3-LICL方法分离的总RNA比试剂盒方法更加最佳。在627bp，262bp的627bp，actin基因中扩展的fapyr1和fachs基因，以及146bp中的26s-18s家庭内政基因到所有选定的样品。

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