Minimize your RNA contaminants with the RiboCut Marker

The purification of ribosome-protected fragments (RPFs) represents one of the critical steps of the ribosome profiling (Ribo-Seq) workflow. RPFs purification is usually carried out via polyacrylamide gel electrophoresis, followed by the excision of fragments with length of ~30 nucleotides. Purification of RPFs has to be performed precisely, as inaccurate excisions can significantly increase the abundance cof ribosomal RNAs (rRNA) and other unwanted contaminants.

The RiboCut Marker consists of three PAGE-purified RNA oligos (27, 29 and 32 nucleotides in length) (Fig. 1) with a phosphate at their 3' end, designed to facilitate the excision of RPFs from denaturing polyacrylamide gels. The RiboCut Marker is compatible with Ribo-Seq samples derived from different species (e.g. human, mouse, rat, C.elegans, D. melanogaster).

 

Available Format

The RiboCut Marker can be used for up to 100 lanes and includes:

Description Quantity Storage
RNA Oligo Mix 1 tube (dry) -80°C 
2 x Loading Dye 1 tube (275 µl) -20°C 

 

 

Figure 1. RiboCut Marker (left lane) and RNA digested (right lane) on a 15% polyacrylamide denaturing gel. The orange rectangle correspond to the gel area where RPFs are found.

Other Ribosome Profiling Products

Precise Monosome Purification with siFractor

During the ribosome profiling protocol, all steps are critical. One of the first steps is the preparation of mono- and polysomal ribosomes. This process includes a sucrose gradient centrifugation step and subsequent fractionation. siTOOLs has developed a device called siFractor which enables easy and reproducible fractionation of mono- and polysomal ribosomes from thin-wall tubes.

Any Species Ribodepletion with Ribo-Seq riboPOOLs

For sequencing RPFs, it is critical to focus on RNA fragments derived from mRNAs rather than non-coding RNAs (ncRNAs) such as ribosomal RNAs (rRNAs). Ribo-Seq riboPOOLs, are specialized ribodepletion kits designed for use prior to RPFs sequencing. The Ribo-Seq riboPOOLs are species-specific (human, mouse, rat, C. elegans, D. melanogaster, L. mexicana, T. brucei, and more) and can be designed for any species with annotated rRNA sequences.