Immagina Biotechnology RiboLace

Improving ribosome profiling

IMMAGINA’s proprietary RiboLace technology enables fast and selective extraction of ribosomes in active translation (both in vitro and in vivo) and purification of ribosome protected fragments (RPFs) for continuing with the ribosome profiling library preparation.

The technology combines a module for sample preparation, three different options for ribosome profiling library preparation/sequencing (RiboLace module 2, LACEseq and circAID-p-seq.) and a kit for gel extraction. The technology allows the user to perform ribosome profiling experiments with a short and simple workflow, no ultracentrifugation steps and low cell input requirements (>100.000 cells in active translation).

Applications

  • Ribosomal profiling
  • Translatome analysis
  • RIBOseq

Key benefits

RiboLace is the only available commercial system to capture native ribosomes in active translation allowing to generate highly accurate translatome data.

It is particularly useful for reliable ribosome profiling with challenging biological samples and with low-input material.

  • Lower Input Requirements

    30-40x lower than standard Riboseq methods

  • Strong Enrichment

    of translated transcripts, which are functionally relevant for biological pathways of interest

  • Reducing Lab Time

    Workflow improvements significantly reducing lab time and allowing for higher sample throughput

  • Improving the Data

    Capturing the active ribosomes is improving greatly the data to noise ratio and increasing the concordance of transcriptomics data with the actual proteome

  • Run Multiple Samples

    Based on beads separation: possibility to run multiple samples in parallel for automated high-throughput (HT) experiments

Ribosome profiling diagram

Active Ribosome Profiling with RiboLace: high quality sequencing data

The possibility to perform Ribo profiling with RiboLace was evaluated on different mammalian cell lines and mouse tissues. As for canonical Ribo-seq, an enrichment of signal from mapped reads along coding sequence regions was observed (Fig. 5A), demonstrating that RiboLace is indeed able to capture bona fide ribosomes. Occupancy meta-profiles, derived from the aggregation of signals on single genes, presented the typical trinucleotide periodicity of the ribosome P-site along coding sequences, which is suggestive of signal derived from ribosomes moving along transcripts (Fig. 5B and 5C).

Products

Ribosome Profiling set image

Ribosome Profiling All-In-One Set

All kits and reagants to perform ribosome profiling from cell lysis to sequencing and data analysis. These products include RiboLace Module 1, LaceSeq, PAGE Extraction Gel and UDI for 9 reactions.

RiboLace Module 1 Kit image

RiboLace Module 1 Kit

Isolate active ribosomes by means of an antibody-free and tag-free pull-down approach. The kit contains all the reagents needed for one-day extraction of ribosomes in active translation.

RiboLace Module 2 photo

RiboLace Module 2 Kit

Prepare ribosome profiling libraries starting from ribosome protected fragments (RPFs) isolated using RiboLace or other protocols. The kit contains buffers, filters and vials, molecular weight ladders for PAGE purification and gel extraction, and adaptors and oligos for library preparation. It includes control RNAs that can be used for RPF size selection. Reverse indexes are not included.

Resources