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High Yield T7 RNA Synthesis Kit

  • Cat.No.ON-040
  • Cas.No.
  • 来源N/A
  • 纯度N/A
  • 储存条件-20℃
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产品描述:The High Yield T7 RNA Synthesis Kit与传统的体外转录反应相比,有着很高的合成量,能够多达25倍,每个反应可以合成高达180μg的RNA。

试剂盒中提供的原料:包括50个反应,20μL/反应。

Component50 rxn kitsStorage
5× Reaction Buffer*200μL-20℃
100mM ATP Solution100μL-20℃
100mM GTP Solution100μL-20℃
100mM UTP Solution100μL-20℃
100mM CTP Solution100μL-20℃
Enzyme Mix75μL-20℃
Control Template(0.5μg/μL)10μL-20℃
DNase I (RNase-free, 1U/μL)50μL-20℃
Nuclease-free H2O1mL-20℃
Ammonium Acetate Stop Solution1.5mL-20℃
Lithium Chloride Precipitation Solution1.5mL-20℃
Gel Loading Buffer0.5mL-20℃


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注:5× Reaction Buffer储存在-70℃,可能会产生白色沉淀,如果有白色沉淀产生,将试剂管加热至37℃ 5min 并且充分混合使其溶解后使用。

1. 按下表中的量完成20μL反应,可根据需要按照比例放大或者减小

ComponentAmount
Nuclease-free H2Oto 20μL
100mM ATP Solution2μL
100mM GTP Solution2μL
100mM CTP solution2μL
100mM UTP solution2μL
Template DNA1μg*
5× Reaction Buffer4μL
Enzynme Mix1.5μL

*转录产量取决于模板的添加量。

2. 充分混合,37℃反应 2h。

3.(可选择)加入1μL DNase I(RNase-free),充分混合,37℃反应30min。

储藏温度:-20℃

质控检测:All components are tested in a functional assay as described in this procedure. A 20-μL reaction containing 1 μg of the control template DNA which codes for a ~800b transcript synthesized >150 μg of RNA after a 2 hr incubation.

1. Milligan JF, Groebe DR, Witherell GW, and Uhlenbeck OC (1987) Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA template. Nucl. Acids Res. 15: 8783–8798.

2. Molecular Cloning, A Laboratory Manual, 2nd edition. (1989) editor C Nolan, Cold Spring Harbor Laboratory Press.


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