DuraScribe®T7转录试剂盒可生成抗RNase A降解的DuraScript®RNA。这是通过在DuraScribe体外转录反应中用2'-氟-dCTP(2'-F-dCTP)和2'-氟-dUTP(2'-F-dUTP)完全替换经典CTP和UTP来实现的。该试剂盒采用uraScribe T7 RNA聚合酶,一种突变型T7 RNA聚合酶,可有效地将2'-F-dCTP和2'-F-dUTP,ATP和GTP整合到RNA中。DuraScribe T7 RNA聚合酶使用与标准T7 RNA聚合酶相同的转录启动子序列。The DuraScribe® T7 Transcription Kit produces DuraScript® RNA that is resistant to RNase A degradation. This is accomplished by completely replacing the canonical CTP and UTP with 2′-Fluorine-dCTP (2′-F-dCTP) and 2′-Fluorine-dUTP (2′-F-dUTP) in the DuraScribe in vitro transcription reaction. The kit features DuraScribe T7 RNA Polymerase, a mutant T7 RNA Polymerase, that efficiently incorporates 2′-F-dCTP and 2′-F-dUTP, ATP and GTP into RNA. DuraScribe T7 RNA Polymerase utilizes the same transcription promoter sequences as standard T7 RNA Polymerase.
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优点: |
稳定:由该试剂盒生产的DuraScript®RNA完全耐受RNase A降解;
高产量:根据转录本的大小,至少产生110~307 pmol的DuraScript®RNA;
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组成成分: |
? DuraScribe T7 Enzyme Mix (200 U/μL): -20°C? DuraScribe T7 10X Reaction Buffer:-20°C? ATP (50 mM): -20°C? GTP (50 mM):-20°C? GTP (50 mM): -20°C? 2′-F-dCTP (50 mM): -20°C? 2′-F-dUTP (50 mM): -20°C? DTT (100 mM): -20°C? RNase-Free DNase I (1 U/μL):-20°C? Control Template DNA (0.5 μg/μL): -20°C |
参考文献: |
1. Molecular Cloning - A Laboratory Manual, Third Edition, 2001. CSHL Press. pp 7.27 – 7.34. J. Sambrook and D. Russell. |