产品详细描述

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.
Components
·         RevertAid™ Reverse Transcriptase
·         RiboLock™ RNase Inhibitor
·         5X Reaction Buffer
·         dNTP Mix,10 mM each
·         Oligo(dT)₁₈ Primer
·         Random Hexamer Primer
·         Control GAPDH RNA
·         Forward GAPDH Primer, 10 µM (5-CAAGGTCATCCATGACAACTTTG-3)
·         Reverse GAPDH Primer, 10 µM (5-GTCCACCACCCTGTTGCTGTAG-3)
·         Water, nuclease free
·         Detailed Protocol
反转录试剂盒
产品信息
全长的第一链cDNA产量高，最长可合成13 kb的cDNA。
增加的反应温度范围：42-50°C。
配套提供RiboLock™ RNase Inhibitor。
完全一试剂盒提供oligo(dT)18和随机六聚体引物。
应用  
第一链cDNA合成，作为RT-PCR和实时RT-PCR模板(1, 2)。
构建全长cDNA文库。
制备杂交用探针。
aRNA合成。 
说明
RevertAid™ First Strand cDNA Synthesis Kit是一个完整的以RNA为模板高效合成第一链cDNA的反应体系。该试剂盒可合成长达13 kb的cDNA。
该试剂盒使用的RevertAidReverse Transcriptase具有的RNase H 活性比AMV reverse transcriptase低。
试剂盒配套提供的重组RiboLock™ RNase Inhibitor有效保护RNA模板免于降解。该产品与逆转录反应完全兼容，因为其在50°C时仍然保持活性。
试剂盒配套提供oligo(dT)18和随机六聚体引物。oligo(dT)18与mRNA 的poly(A)尾选择性结合。随机六聚体引物转录无需poly(A) 尾，因此可转录mRNA的5’-端区域，也可以无poly(A) 尾的RNA (micro RNAs)为模板合成cDNA。试剂盒也可使用基因特异性引物。
第一链cDNA可直接作为PCR (见RT-PCR操作说明书)、实时PCR或第二链cDNA 合成(见操作说明书)的模板。 
质量控制
功能测试：以多聚腺苷酸RNA转录子为模板合成第一链cDNA(分别测试特异性引物、oligo(dT)18和随机六聚体引物)。合成的cDNA作为后续PCR的模板。 
组分
RevertAid™ Reverse Transcriptase
RiboLock™ RNase Inhibitor
5X Reaction Buffer
dNTP Mix,10 mM each
Oligo(dT)18 Primer
Random Hexamer Primer
Control GAPDH RNA
Forward GAPDH Primer, 10 µM (5-CAAGGTCATCCATGACAACTTTG-3)
Reverse GAPDH Primer, 10 µM (5-GTCCACCACCCTGTTGCTGTAG-3)
Water, nuclease free
保存与运输： -20°C