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fermentas逆轉(zhuǎn)錄試劑盒RevertAid First Strand cDNA Synthesis Kit說(shuō)明書(shū)

Thermo Scientific RevertAid First Strand cDNA Synthesis Kit是一套用于以RNA為模板高效合成*鏈cDNA的完整系統(tǒng)，可合成長(zhǎng)達(dá)13kb的cDNA。試劑盒配套提供的RiboLockRNase Inhibitor，可有效防止RNA模板的降解。試劑盒配套提供oligo(dT)18引物和隨機(jī)六聚體引物。oligo(dT)18引物可以選擇性地與mRNA中的poly (A)尾結(jié)合。隨機(jī)六聚體引物無(wú)需poly (A)尾，因此可轉(zhuǎn)錄mRNA 5’-端區(qū)域，也可以無(wú)poly (A)尾的RNA (如micoRNA)為模板合成cDNA。該試劑盒也可使用基因特異性引物。

優(yōu)點(diǎn)：

• 合成全長(zhǎng)的*鏈cDNA，zui長(zhǎng)可合成13kb的cDNA
• *反應(yīng)溫度為42℃
• *一提供RT反應(yīng)所需的所有組分

應(yīng)用:

• *鏈cDNA合成，作為RT-PCR和實(shí)時(shí)RT-qPCR的模板
• 構(gòu)建全長(zhǎng)cDNA文庫(kù)
• 反義RNA合成

fermentas逆轉(zhuǎn)錄試劑盒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合成全長(zhǎng)cDNA
1 μg小鼠心臟總RNA作為模板，用oligo(dT)18 引物進(jìn)行逆轉(zhuǎn)錄反應(yīng)。由此合成的cDNA，再作為T(mén)aq DNA Polymerase后續(xù)PCR擴(kuò)增的模板。末端2024 bp片段的成功擴(kuò)增，表明長(zhǎng)度為13.8 kb的全長(zhǎng)RNA片段得到成功的逆轉(zhuǎn)錄。

產(chǎn)品組成
RevertAid First Strand cDNA Synthesis Kit包含RevertAid Reverse Transcriptase、RiboLock RNase Inhibitor、5x Reaction Buffer、dNTP Mix、Oligo(dT)18 引物、隨機(jī)六聚體引物、對(duì)照GAPDH RNA、10 μM 正向GAPDH引物、10 μM反向GAPDH 引物以及nuclease-free water。