| 锦鲤保护性组织RNA提取及引物扩增研究 |
| The Research of RNA Extraction and Primer Amplification from Protective Tissues of the Koi (Cyprinus carpio) |
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| DOI： |
| 中文关键词:锦鲤 保护性组织 RNA提取 引物扩增 |
| 英文关键词:ornamental carp protective tissues RNA extraction primer amplification |
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| 中文摘要: |
| 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TR izo l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( B- 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NCB I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。
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| 英文摘要: |
| The integrity of RNA extracted and purified from various organizations is a key step for molecular biology research to genes of koi in growth and development, stress - express and variation of body color. With reducing harm to precious individual in the maximal degree, The protective tissues of fins, scales, gills and blood can be researched for the expression of genes at the same time. Total RNA was extracted by means of TRizol, these RNA have complete fragment and high purity. The results showed that the CDNA reversed from RNA of these organiza- tions have high quality. The PCR obtained distinct bands amplified by the primers of β - actin, hsp70 and MT, the highest comparability 95% is show when these sequences are compared using BLAST aligning on NCBI. In summary, this method can fully protect the individual integrity of Koi, which would benefical for the further research of molecular biology. |
| 王亚军,刘必谦,吴立山,洪松柏.2008.锦鲤保护性组织RNA提取及引物扩增研究[J].水生态学杂志,29(2):116-119. |
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