m6A sites can be identified with eCLIP
- N6-methyladenosine (m6A) is an abundant modification in mRNA.
- m6A has diverse molecular functions- modulates many mRNA processes, as mRNA export, stability, translation efficiency, etc.
- m6A-eCLIP is a modified version of eCLIP (Van Nostrand et al. Nature Methods, 2016) using a validated m6A antibody.
- m6A-eCLIP uses 40-fold less mRNA and requires fewer PCR cycles than previously published methods.
- m6A-eCLIP captures m6A motif and enrichment at stop codons of genes.
Rigorous in-line validation
1.Library yield and size distribution confirmed by Tapestation and qPCR
2.Generate complex libraries with high yields.
3.Sequencing to yield 20 million reads
4.Identify m6a sites with a single nucleoside resolution.
RNA-seq control significantly improves signal/noise to identify true modification targets