IN VITRO TRANSCRIPTION (RNA SYNTHESIS)
KIT FOR HIGH-YIELD SYNTHESIS OF RNA IN VITRO
RNA体外高效合成试剂盒
The principle of operation of the kit is based on the enzymatic synthesis of RNA molecules on a DNA template using DNA-dependent RNA polymerase of bacteriophage T7. The kit
contains all the necessary reactive chemicals for obtaining a high yield of RNA transcripts in a minimal reaction time: T7 RNA polymerase, rNTP mixture, (× 5) buffer for T7 transcription, (× 25) DTT, sterile water
试剂盒的工作原理是利用噬菌体T7的DNA依赖性RNA聚合酶在DNA模板上酶促合成RNA分子。工具包
包含在最短反应时间内获得高产量RNA转录物所需的所有反应性化学物质：T7 RNA聚合酶、rNTP混合物、（×5）T7转录缓冲液、（×25）DTT、无菌水
T7 RNA POLYMERASE
T7 RNA聚合酶
T7 RNA Polymerase is a DNA-dependent RNA polymerase with strict specificity for its respective double-stranded promoters. It exhibits extremely high specificity for its cognate promoter sequences. Only T7 DNA or DNA cloned downstream from a T7 promoter can serve as a template for T7 RNA Polymerase-directed RNA synthesis.
T7 RNA聚合酶是一种DNA依赖性RNA聚合酶，对其各自的双链启动子具有严格的特异性。它对同源启动子序列表现出极高的特异性。只有T7 DNA或从T7启动子下游克隆的DNA可以作为T7 RNA聚合酶定向RNA合成的模板。
It catalyzes the 5’→3’ synthesis of RNA on either single-stranded DNA or double-stranded DNA downstream from its promoter. 它催化5'→3’在其启动子下游的单链DNA或双链DNA上合成RNA。
Feature:
Incorporates modified nucleotides (e.g., aminoallyl-, biotin-, fluorescein-, digoxigenin- labeled nucleotides) 结合修饰核苷酸（例如，氨基烯丙基、生物素、荧光素、地高辛标记的核苷酸）
Applications:
Synthesis of unlabeled and labeled RNA 未标记和标记RNA的合成
Consensus promoter sequence:
T7: TAATACGACTCACTATAGGGAGA
一致启动子序列：T7:Taatacgactcacatagggaga
QC Tests:
Activity, SDS-PAGE/purity, DNase, RNase, endonuclease, transcription.
活性，SDS-PAGE/纯度，DNA酶，RNA酶，核酸内切酶，转录。
Source:
Recombinant E. coli strain. 重组大肠杆菌菌株。
Storage Buffer:
50mM Tris-HCl (pH 7.5 at 25°C), 1mM EDTA, 20mM 2-mercaptoethanol, 100mM NaCI, 0.1% (v/v) Triton® X-100 and 50% (v/v) glycerol.
50mM Tris-HCl（25°C时pH 7.5）、1mM EDTA、20mM 2-巯基乙醇、100mM NaCl、0.1%（v/v）Triton®X-100和50%（v/v）甘油。
Storage Conditions:
Store at –20°C. 储存条件：储存温度为–20°C。
Unit Definition:
One unit is defined as the amount of enzyme required to catalyze the incorporation of 1 nmol of NTP into acid-insoluble product in 60 minutes at 37°C in a total volume of 100μl. The reaction conditions are: 50mM Tris-HCl (pH 7.5 at 25°C), 6 mM MgCl2,10 mM DTT, 2mM spermidine, 0.5mM each of ATP, GTP, CTP, and UTP, 0.5μCi [3H]CTP and 2μg DNA.
一个单位被定义为在37°C下60分钟内催化1 nmol NTP并入酸不溶性产物所需的酶量，总体积为100μl。反应条件为：50mM Tris-HCl（25°C下pH 7.5），6 mM MgCl2，10 mM DTT，2mM亚精胺，ATP、GTP、CTP和UTP各0.5mM，0.5μCi[3H]CTP和2μg DNA。
