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100次
Thermo Scientific RevertAid第一链cDNA合成试剂盒配套提供oligo(dT)18引物和随机六聚体引物。oligo(dT)18引物可以选择性地与mRNA中的poly (A)尾结合。随机六聚体引物无需poly (A)尾,因此可转录mRNA 5’-端区域,也可以无poly (A)尾的RNA (如micoRNA)为模板合成cDNA。该试剂盒也可使用基因特异性引物。
特点:
• 可以高效合成长达13kb的全长第一链cDNA
• 最佳活性温度为42°C
• 完全一提供RT反应所需的所有组分
应用
• 第一链cDNA合成,作为RT-PCR和实时RT-qPCR的模板
• 构建全长cDNA文库
• 反义RNA合成
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文献和实验Protocol for First-strand cDNA Synthesis
℃for 10 minutes and then at 37℃for 60 minutes.Stop the reaction by heating at 70℃for 10 minutes. Chill on ice.NoteThe synthesized cDNA can be amplified by the PCR (see Protocols for PCR using Taq and Pfu DNA Polymerases) without intermediate phenol
of mRNA) or random hexamers; the kit used here contains both kinds); nucleotides for DNA synthesis (dNTPs); MgC2 and buffers required by the enzyme, which are all present in the 5x Reaction Mix. Materials * iScript cDNA Synthesis Kit (BIORAD
methylation analysis, PCR, and multiplex PCR. The purified total RNA is ready to use in any application, such as RT-PCR, real-time RT-PCR, differential display, cDNA synthesis, Northern, dot, and slot blot analyses, and microarray analysis. The kits form
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