产品信息

• 产品组分信息

  本产品提供以下试剂或组分：

  NEB #	名称	组分货号	储存温度	数量	浓度
  M0458S     -20       RtcB Ligase M0458SVIAL -20 1 x 0.025 ml 15 µM   RtcB Reaction Buffer B0790AVIAL -20 1 x 0.5 ml 10 X   MnCl2 B1761SVIAL -20 1 x 1 ml 10 X   GTP N2080AVIAL -20 1 x 0.05 ml 10 mM

• 特性和用法

  反应条件

  1X RtcB Reaction Buffer
  Supplement with 0.1 mM GTP 和 1 mM MnCl₂
  Incubate at 37°C

  1X RtcB Reaction Buffer
  50 mM Tris-HCl
  75 mM KCl
  3 mM MgCl₂
  10 mM DTT
  (pH 8.3 @ 25°C)

  贮存溶液

  50 mM NaCl
  10 mM Tris-HCl
  0.1 mM EDTA
  1 mM DTT
  50% Glycerol
  pH 7.4 @ 25°C

• 优势和特性

  应用特性

  • Ligate ssRNA or ssDNA with a 3´-phosphate or a 2´,3´-cyclic phosphate to the 5´-OH of ssRNA
  • Circularization of ssRNA with compatible ends, as above
  • Detection of 3´-phosphorylated or 5´ -OH RNA using labeled probes

• 注意事项

  1. 对于困难底物，添加 15% 的 PEG 8000 可以提高连接的产量。

• 参考文献

  1. Naoko, T., Shuman, S. (2011). J. Biol. Chem. 286, 7727-7731.
  2. Naoko, T. et al. (2011). J. Biol. Chem. 286, 43134-43143.
  3. Chakravarty, A. K. et al (2012). Proc. Natl. Acad. Sci. USA. 109, 6072-6077.

操作说明、说明书 & 用法

• 操作说明

  1. Typical Protocol for RNA Ligation (NEB #M0458)

工具 & 资源

• 选择指南

  • Properties of DNA and RNA Ligases
  • RNA Ligase Selection Chart
  • Substrate-based Ligase Selection Chart

FAQs & 问题解决指南

• FAQs

  1. Does secondary structure affect ligation by RtcB Ligase?
  2. Can RtcB Ligase ligate DNA?
  3. What is the optimum reaction temperature for RtcB Ligase?
  4. Can RtcB Ligase be used in other NEBuffers?
  5. How can I increase ligation efficiency?
  6. Can magnesium, or another divalent cation, be substituted for manganese?
  7. Does PEG improve ligation efficiency?
  8. Can I purchase large amounts of an existing Enzyme for Innovation?
  9. What are Enzymes for Innovation?