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品名
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規(guī)格
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RevertAid™ First Strand cDNA Synthesis Kit
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K1622
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Fermentas（MBI）
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100 react
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¥1871
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反轉(zhuǎn)錄試劑盒
全長的第一鏈cDNA產(chǎn)量高，最長可合成13 kb的cDNA。
增加的反應(yīng)溫度范圍：42-50°C。
配套提供RiboLock™ RNase Inhibitor。
完全一試劑盒提供oligo(dT)18和隨機(jī)六聚體引物。
應(yīng)用
第一鏈cDNA合成，作為RT-PCR和實(shí)時(shí)RT-PCR模板(1, 2)。
構(gòu)建全長cDNA文庫。
制備雜交用探針。
aRNA合成。
說明
RevertAid™ First Strand cDNA Synthesis Kit是一個(gè)完整的以RNA為模板高效合成第一鏈cDNA的反應(yīng)體系。該試劑盒可合成長達(dá)13 kb的cDNA。
該試劑盒使用的RevertAidReverse Transcriptase具有的RNase H 活性比AMV reverse transcriptase低。
試劑盒配套提供的重組RiboLock™ RNase Inhibitor有效保護(hù)RNA模板免于降解。該產(chǎn)品與逆轉(zhuǎn)錄反應(yīng)完全兼容，因?yàn)槠湓?0°C時(shí)仍然保持活性。
試劑盒配套提供oligo(dT)18和隨機(jī)六聚體引物。oligo(dT)18與mRNA 的poly(A)尾選擇性結(jié)合。隨機(jī)六聚體引物轉(zhuǎn)錄無需poly(A) 尾，因此可轉(zhuǎn)錄mRNA的5’-端區(qū)域，也可以無poly(A) 尾的RNA (micro RNAs)為模板合成cDNA。試劑盒也可使用基因特異性引物。
第一鏈cDNA可直接作為PCR (見RT-PCR操作說明書)、實(shí)時(shí)PCR或第二鏈cDNA 合成(見操作說明書)的模板。
質(zhì)量控制
功能測試：以多聚腺苷酸RNA轉(zhuǎn)錄子為模板合成第一鏈cDNA(分別測試特異性引物、oligo(dT)18和隨機(jī)六聚體引物)。合成的cDNA作為后續(xù)PCR的模板。
Features
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Full-length first strand cDNA up to 13 kb.
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Increased reaction temperatures in the range of 42-50°C.
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Supplied with the recombinant RiboLock™ RNase Inhibitor.
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Complete – oligo(dT)18 and random hexamer primers included with the kit.
Applications
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First strand cDNA synthesis for RT-PCR and real-time RT-PCR (1, 2).
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Construction of full length cDNA libraries.
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Generation of probes for hybridization.
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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
北京
