fermentas逆转录试剂盒RevertAid First Strand cDNA Synthesis Kit说明书

Thermo Scientific RevertAid First Strand cDNA Synthesis Kit是一套用于以RNA为模板高效合成*链cDNA的完整系统，可合成长达13kb的cDNA。试剂盒配套提供的RiboLockRNase Inhibitor，可有效防止RNA模板的降解。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。

优点：

• 合成全长的*链cDNA，zui长可合成13kb的cDNA
• *反应温度为42℃
• *一提供RT反应所需的所有组分

应用:

• *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

fermentas逆转录试剂盒RevertAid First Strand cDNA Synthesis Kit 

RevertAid First Strand cDNA Synthesis Kit
Features
·         Full-length first strand cDNA up to 13 kb.
·         Increased reaction temperatures in the range of 42-50°C.
·         Supplied with the recombinant RiboLock? RNase Inhibitor.
·         Complete – oligo(dT)18 and random hexamer primers included with the kit.
Applications
·         First strand cDNA synthesis for RT-PCR and real-time RT-PCR (1, 2).
·         Construction of full length cDNA libraries.
·         Generation of probes for hybridization.
·         aRNA synthesis.
Description
The RevertAid? First Strand cDNA Synthesis Kit is a complete system for efficient synthesis of first strand cDNA from RNA templates. The kit is suitable for synthesis of cDNA up to 13 kb.
The kit uses RevertAid? Reverse Transcriptase which has lower RNase H activity, compared to AMV reverse transcriptase.
The recombinant RiboLock? RNase Inhibitor, supplied with the kit, effectively protects RNA template from degradation. It is fully compatible with reverse transcription reaction, as it maintains activity at temperatures up to 55°C.
The kit is supplied with both oligo(dT)18 and random hexamer primers. The oligo(dT)18 anneals selectively on the poly(A) tail of mRNA. Random hexamer primers do not require the presence of poly(A). Therefore, they can be used for transcription of the 5-end regions of mRNA or cDNA synthesis using RNA without poly(A) tail e.g. micro RNAs. Gene-specific primers may also be used with the kits.
The first strand of cDNA can be directly used as a template in PCR (see RT-PCR protocol), real-time PCR or in second strand cDNA synthesis (see protocol). 
Quality Control
The kit is functionally tested in RT-PCR using 100 fg control GAPDH RNA and GAPDH control primers generated a 496 bp product visible on agarose gel after ethidium bromide staining.

is Kit

RevertAid First Strand cDNA Synthesis Kit合成全长cDNA
1 μg小鼠心脏总RNA作为模板，用oligo(dT)18 引物进行逆转录反应。由此合成的cDNA，再作为Taq DNA Polymerase后续PCR扩增的模板。末端2024 bp片段的成功扩增，表明长度为13.8 kb的全长RNA片段得到成功的逆转录。

产品组成
RevertAid First Strand cDNA Synthesis Kit包含RevertAid Reverse Transcriptase、RiboLock RNase Inhibitor、5x Reaction Buffer、dNTP Mix、Oligo(dT)18 引物、随机六聚体引物、对照GAPDH RNA、10 μM 正向GAPDH引物、10 μM反向GAPDH 引物以及nuclease-free water。