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| 品名 | 货号 | 品牌 | 规格 | 售价 |
| RevertAid™ First Strand cDNA Synthesis Kit | K1622 | Fermentas（MBI） | 100 react | 询价 |
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
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