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从养殖锦鲤各种组织中提取完整的RNA,是分子生物学研究锦鲤应激表达、体色变异、生长发育等功能基因表达的关键实验。在不伤害养殖锦鲤个体健康的情况下,取用鱼体的鳍条、鳞片、鳃丝、血液4种保护性组织,可以最大程度地减少对珍贵锦鲤个体的损伤,并达到研究相关基因表达的目的。本研究利用TRizol法提取RNA,获得的各样品RNA条带完整、纯度高。结果表明,4种保护性组织提取的RNA经过逆转录反应,得到了高质量的CDNA。内标基因(β-actin)、热激蛋白基因(hsp70)、金属硫蛋白基因(MT)扩增得到清晰的条带,通过NCBI数据上的BLAST比对证明,各序列的同源性在95%以上。利用本研究的方法,可以充分保护养殖锦鲤个体的完好性,用于进一步的分子生物学研究。
The extraction of intact RNA from various tissues of cultured Koi is a key experiment in molecular biology to study the expression of functional genes such as stress, body color, growth and development of Koi. Without damaging the individual health of cultured Koi, taking four kinds of protective tissues, such as the fin, scales, gill filaments and blood of the fish body, can minimize the damage to the precious Koi individual and achieve the research related gene The purpose of expression. In this study, TRizol method was used to extract RNA, and each sample obtained was complete with high purity. The results showed that RNA extracted from the four kinds of protective tissues obtained high quality CDNA by reverse transcription reaction. A clear band was obtained by amplification of β-actin, hsp70 and MT, and the BLAST alignment of NCBI data showed that the homology of each sequence was 95 %the above. The method of this study can fully protect the integrity of breeding Koi individuals, for further molecular biology.