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表1:人源let系列miRNA及其miR-98序列
| miRNA ID | miRNA SEQ |
| hsa-let-7a | UGAGGUAGUAGGUUGUAUAGUU |
| hsa-let-7b | UGAGGUAGUAGGUUGUGUGGUU |
| hsa-let-7c | UGAGGUAGUAGGUUGUAUGGUU |
| hsa-let-7d | AGAGGUAGUAGGUUGCAUAGUU |
| hsa-let-7e | UGAGGUAGGAGGUUGUAUAGUU |
| hsa-let-7f | UGAGGUAGUAGAUUGUAUAGUU |
| hsa-let-7g | UGAGGUAGUAGUUUGUACAGUU |
| hsa-let-7i | UGAGGUAGUAGUUUGUGCUGUU |
| hsa-miR-98 | UGAGGUAGUAAGUUGUAUUGUU |
以let-7a为参考,其它let系列miRNA与其比对后的碱基差异(以红色标记)
表2:通过各miRNA特异引物分别检测人源let系列miRNA来进行检测分辨率测试
| miRNA qPCR Detection System的相对检测分辨率 | |||||||||
| miRNA 质粒模板 |
All-in-One™ miRNA qPCR Primer | ||||||||
| let-7a | let-7b | let-7c | let-7d | let-7e | let-7f | let-7g | let-7i | miR-98 | |
| hsa-let-7a | 100.00% | 0.01% | 0.33% | 0.08% | 3.21% | 0.41% | 0.00% | 0.00% | 0.00% |
| hsa-let-7b | 0.00% | 100.00% | 0.42% | 0.00% | 0.00% | 0.00% | 0.00% | 0.00% | 0.00% |
| hsa-let-7c | 0.32% | 2.05% | 100.00% | 0.00% | 0.00% | 0.00% | 0.00% | 0.00% | 0.00% |
| hsa-let-7d | 8.97% | 0.00% | 0.00% | 100.00% | 0.00% | 0.00% | 0.00% | 0.00% | 0.00% |
| hsa-let-7e | 1.51% | 0.00% | 0.01% | 0.00% | 100.00% | 0.01% | 0.00% | 0.00% | 0.00% |
| hsa-let-7f | 2.67% | 0.00% | 0.01% | 0.00% | 0.04% | 100.00% | 0.00% | 0.00% | 0.00% |
| hsa-let-7g | 0.17% | 0.00% | 0.00% | 0.00% | 0.00% | 0.04% | 100.00% | 0.00% | 0.00% |
| hsa-let-7i | 0.00% | 0.00% | 0.00% | 0.00% | 0.00% | 0.00% | 0.01% | 100.00% | 0.00% |
| hsa-miR-98 | 0.01% | 0.00% | 0.00% | 0.00% | 0.00% | 0.00% | 0.00% | 0.00% | 100.00% |
克隆let系列的miRNA,各miRNA特异引物通过All-in-One™ miRNA qPCR detection Kit分别检测以105分子数的此miRNA质粒。以引物与miRNA完全匹配所得的Ct值为对照,各碱基有差异的miRNA 的Ct值与之相比,通过公式(相对分辨率=2-ΔCt ×100%)即可得到miRNA qPCR Detection System 的检测分辨率。表中标记为红色的为引物和miRNA序列完全匹配的检测分辨率。灰色的为单碱基差异的检测分辨率,黄色的为双碱基差异的分辨率,白色的为2个以上碱基差异的检测分辨率
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