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Home Page |MicroRNA Research |Discovery & Cloning

Discovery & Cloning

Immunopurify microRNA and piRNA complexes and their associated RNAs

Discovery & Cloning Systems

  • Immunoprecipitate microRNA factors
  • Immunoprecipitate piRNA factors
  • Clone microRNAs with the Global MicroRNA Amplification Kit

Immunopurify microRNAs & piRNAs and associated complexes

Use epitope-tagged microRNA processing factors and PIWI factors to pull-down the proteins and their associated RNAs.
1.  Transfect or transduce the miR-Snare or PIWI-Snare factor
2.  Immunopurify the protein factor
3.  Enrich and discover new small RNAs and protein complexes
4.  Identify messenger RNAs driven to RISC complexes

 

Clone Mature MicroRNAs

SBI’s Small RNA Amplification and Cloning system includes 3 steps:
1.  Ligation of an adaptor to the 3’-end of the RNA;
2.  Reverse transcription of the RNA and attachment of a 5’-end adaptor in the same reaction by template switching;
3.  PCR amplification of the cDNA with a PCR polymerase mixture that incorporates a proof reading function.

The amplified cDNA is ready for qPCR studies with no purification necessary. For microarray studies, the amplified cDNA is first purified on spin column and labeled via T7 in vitro transcription in the presence of fluorescently labeled nucleotides.