ICE
Inosine Chemical Erasing
ICE (Sakurai et al., 2010) (Sakurai et al., 2014) (Suzuki et al., 2015) followed by NGS identifies adenosine-to-inosine editing. In this method, RNA is treated with acrylonitrile, while control RNA is untreated. Control and treated RNAs are reverse-transcribed and PCR-amplified. Inosines in RNA fragments treated with acrylonitrile cannot be reverse-transcribed. Deep sequencing of the cDNA prepared from control and treated RNA provides high-resolution reads of inosines in RNA fragments.
Advantages:
- Provides mapping of adenosine-to-inosine editing
- Can be performed with limited material
Disadvantages:
- Nonlinear PCR amplification can lead to biases, affecting reproducibility
- Amplification errors caused by polymerases will be represented and sequenced incorrectly
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
Ramaswami G. and Li J. B. Identification of human RNA editing sites: A historical perspective. Methods. 2016;107:42-47
Frye M., Jaffrey S. R., Pan T., Rechavi G. and Suzuki T. RNA modifications: what have we learned and where are we headed? Nat Rev Genet. 2016;17:365-372
References:
Ishida K., Miyauchi K., Kimura Y., et al. Regulation of gene expression via retrotransposon insertions and the noncoding RNA 4.5S RNA. Genes Cells. 2015;
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History: ICE
Revision by sbrumpton on 2017-06-21 07:50:22 - Show/Hide
Inosine Chemical Erasing
ICE (Sakurai et al., 2010) (Sakurai et al., 2014) (Suzuki et al., 2015) followed by NGS identifies adenosine-to-inosine editing. In this method, RNA is treated with acrylonitrile, while control RNA is untreated. Control and treated RNAs are reverse-transcribed and PCR-amplified. Inosines in RNA fragments treated with acrylonitrile cannot be reverse-transcribed. Deep sequencing of the cDNA prepared from control and treated RNA provides high-resolution reads of inosines in RNA fragments.
Advantages:- Provides mapping of adenosine-to-inosine editing
- Can be performed with limited material
Disadvantages:- Nonlinear PCR amplification can lead to biases, affecting reproducibility
- Amplification errors caused by polymerases will be represented and sequenced incorrectly
Reagents:Illumina Library prep and Array Kit SelectorReviews:Ramaswami G. and Li J. B. Identification of human RNA editing sites: A historical perspective. Methods. 2016;107:42-47Frye M., Jaffrey S. R., Pan T., Rechavi G. and Suzuki T. RNA modifications: what have we learned and where are we headed? Nat Rev Genet. 2016;17:365-372References:Ishida K., Miyauchi K., Kimura Y., et al. Regulation of gene expression via retrotransposon insertions and the noncoding RNA 4.5S RNA. Genes Cells. 2015;