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进口试剂

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MessageMuter™ shRNA Production Kit

进口试剂

MessageMuter™ shRNA Production Kit

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产品名称:

MessageMuter™ shRNA Production Kit

货号
MM031110
规格
10 Rxns
存储温度
-20℃
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产品描述

The MessageMuter shRNA Production Process

The MessageMuter shRNA Production Kit utilizes a simple and unique 3-step process (Figure 2) that yields shRNA in about 4 hours. A 17-base T7 Promoter Oligo is annealed to a DNA oligo designed by the user using detailed instructions provided. The ends of the annealed duplex are "filled-in" using the Exo-Minus Klenow to obtain a linear dsDNA with a T7 RNAP promoter at its 5'-end. Following in vitro transcription, the RNA produced spontaneously forms a hairpin structure (shRNA). Following clean-up, the shRNA is ready for transfection into cultured cells. The MessageMuter shRNA kit provides reagents to produce 10 shRNAs in sufficient quantities for hundreds of transfections.

Applications

  • Production of shRNA, micro RNA (miRNA) studies, or other applications.

Benefits

  • Produces shRNA in vitro using one user-designed oligo and one in vitro transcription reaction.
  • No need for annealing of sense and anti-sense RNA strands as required by other in vitro transcription-based methods.

 

   Figure 1. Short hairpin RNA (shRNA) directed against firefly luciferase. The shRNA produced using the MessageMuter™ shRNA Production Kit contains a sequence homologous to the target mRNA (sense sequence), a "loop" region and a sequence complementary to the target sequence (anti-sense sequence).

  Figure 2. Overview of the method used to produce shRNA using the MessageMuter™ shRNA Production Kit. All reagents are supplied except a 60- to 76-base DNA oligo designed by the user as specified in the kit.

References

  1. McManus, M.T. (2002) RNA 8 , 842.
  2. Paddison, P.J. et. al. (2002) Genes & Development 16, 948.
  3. Yu, J-Y et. al. (2002) Proc. Natl. Acad. Sci. USA 99 (9), 6047.
  4. Meis, J.E. (2003) EPICENTRE Forum 10 (2), 1.
  5. Meis, J.E. (2004) EPICENTRE Forum 11 (1), 1.
  6. Lin, Z.P. et. al. (2008) Mol. Pharmacol. 73, 243

Use of dsRNA for RNA interference by for-profit organizations requires a license from the Carnegie Institution of Washington. For-profit institutions should contact Gloria Brienza of the Carnegie Institution of Washington, 1530 P Street, N.W., Washington, D.C. 20005-1910. Email: gbrienza@pst.ciw.edu.

 

 Catalog No. Size

MessageMuter™ shRNA Production Kit
   MM031110 10 Reactions
Contents:
T7 Promoter Oligo, Exo-Minus Klenow DNA Polymerase, 5X Annealing Buffer, dNTPs, AmpliScribe™ T7-Flash™ Enzyme Mix, AmpliScribe™ T7-Flash™ 10X Reaction Buffer, NTPs, DTT, RNase-Free Water, Control Oligo, and RiboGuard™ RNase Inhibitor.

 

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