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
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
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.