Pol II CLIP
Crosslinking and Immunoprecipitation of RNA Polymerase II
Pol II CLIP was developed to isolate and sequence noncoding RNAs involved in transcriptional regulation, particularly RNAPII-mediated transcription (Li et al., 2015). Briefly, cells are UV-crosslinked at 254 nm and lysed. Anti-RNAPII antibodies are used to immunoprecipitate the RNAPII complexes. Next, the RNAPII complexes are separated via proteinase K reverse-crosslinking. The released RNA released is isolated, reverse-transcribed, and prepared into sequencing libraries. The CLIP RNA samples are ready for sequencing.
Advantages:
- Identifies ncRNA sequences involved in transcriptional regulation
- Simple procedure
Disadvantages:
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
Qu S., Yang X., Li X., Wang J., Gao Y., et al. Circular RNA: A new star of noncoding RNAs. Cancer Lett. 2015;365:141-148
References:
Li Z., Huang C., Bao C., et al. Exon-intron circular RNAs regulate transcription in the nucleus. Nat Struct Mol Biol. 2015;
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History: Pol II CLIP
Revision by sbrumpton on 2017-06-21 07:50:24 - Show/Hide
Crosslinking and Immunoprecipitation of RNA Polymerase II
Pol II CLIP was developed to isolate and sequence noncoding RNAs involved in transcriptional regulation, particularly RNAPII-mediated transcription (Li et al., 2015). Briefly, cells are UV-crosslinked at 254 nm and lysed. Anti-RNAPII antibodies are used to immunoprecipitate the RNAPII complexes. Next, the RNAPII complexes are separated via proteinase K reverse-crosslinking. The released RNA released is isolated, reverse-transcribed, and prepared into sequencing libraries. The CLIP RNA samples are ready for sequencing.
Advantages:- Identifies ncRNA sequences involved in transcriptional regulation
- Simple procedure
Disadvantages:Reagents:Illumina Library prep and Array Kit SelectorReviews:Qu S., Yang X., Li X., Wang J., Gao Y., et al. Circular RNA: A new star of noncoding RNAs. Cancer Lett. 2015;365:141-148References:Li Z., Huang C., Bao C., et al. Exon-intron circular RNAs regulate transcription in the nucleus. Nat Struct Mol Biol. 2015;