CIP-TAP
Alkaline Phosphatase, Calf Intestine-Tobacco Acid Pyrophosphatase Sequencing
CIP-TAP maps capped small RNAs (Gu et al., 2012). In this method, RNA is treated with CIP followed by 3′-end linker ligation. Next, the RNA is treated with TAP, followed by 5′-end linker ligation. The fragments are reverse-transcribed to cDNA, PCR-amplified, and sequenced. Deep sequencing provides single-nucleotide resolution reads of the capped small RNAs.
Advantages:
- Identifies capped small RNAs missed by CapSeq
- High throughput
Disadvantages:
- Nonlinear PCR amplification can lead to biases affecting reproducibility
- Amplification errors caused by polymerases
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
No reviews yet
References:
Gu W., Lee H. C., Chaves D., et al. CapSeq and CIP-TAP identify Pol II start sites and reveal capped small RNAs as C. elegans piRNA precursors. Cell. 2012;151:1488-1500
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History: CIP-TAP
Revision by sbrumpton on 2017-06-21 07:50:23 - Show/Hide
Alkaline Phosphatase, Calf Intestine-Tobacco Acid Pyrophosphatase Sequencing
CIP-TAP maps capped small RNAs (Gu et al., 2012). In this method, RNA is treated with CIP followed by 3'-end linker ligation. Next, the RNA is treated with TAP, followed by 5'-end linker ligation. The fragments are reverse-transcribed to cDNA, PCR-amplified, and sequenced. Deep sequencing provides single-nucleotide resolution reads of the capped small RNAs.
Advantages:- Identifies capped small RNAs missed by CapSeq
- High throughput
Disadvantages:- Nonlinear PCR amplification can lead to biases affecting reproducibility
- Amplification errors caused by polymerases
Reagents:Illumina Library prep and Array Kit SelectorReviews:No reviews yet
References:Gu W., Lee H. C., Chaves D., et al. CapSeq and CIP-TAP identify Pol II start sites and reveal capped small RNAs as C. elegans piRNA precursors. Cell. 2012;151:1488-1500