AHARIBO for RNA analysis is dedicated to the analysis of the translatome.
The product is based on an IMMAGINA technology called Minimally Invasive Non-canonical Amino acid Tagging and Isolation of Ribosomes (Ribo-MINATI).
Classical methods for translatome analysis such as polysome profiling or affinity purification-based techniques followed by RNA-seq are characterized labor-intensive protocols and have so far led to relatively poor correlations between mRNA and protein levels. AHARIBO for RNA analysis relies on a fast, low-input protocol and can capture the dynamics of translation across different biological conditions. If combined with AHARIBO for de novo proteome analysis, the kit allows for the parallel isolation and downstream analysis of translated RNAs (by RNA-seq or qPCR) and the associated newly synthesized proteins.
AHARIBO for RNA analysis is based on the incorporation of non-canonical amino acid L-azidohomoalanine (AHA) within ribosome-bound nascent peptide chains, followed by their stabilization on the ribosomes by means of a blocking molecule and the final click chemistry- mediated pulldown of translational complexes through the AHA-tagged nascent polypeptides.
Our core findings show that AHARIBO can enrich for (i) protein-coding RNAs, (ii) ribosome-associated regulatory RNAs, and (iii) translated RNAs annotated as non-coding. AHARIBO-identified translatome correlates better than total RNA with the respective protein levels in the dynamic framework of neuronal differentiation of mouse embryonic stem cells. AHARIBO therefore represents an effective tool to explore quantitative relationships between transcript and protein levels, offering a reliable and accurate approach for taking snapshots of active translation processes.
Shipping: dry ice and +4°C
Storage Conditions: store components as indicated on the data sheet
Shelf Life: 5 months
The kit contains reagents for 6 experiments
Protocol AHARIBO RNA