ribosome-seq方法

Ribosome profiling (also known as ribosome sequencing or Ribo-seq) is a method used to study translation in cells. It involves the capture and sequencing of ribosome-protected mRNA fragments to give insight into the dynamics of protein synthesis.

The basic steps of ribosome profiling are as follows:

1. Cells are grown under conditions where translation is actively occurring.
2. Ribosomes are cross-linked to mRNA using a chemical such as formaldehyde.
3. Cells are lysed and the ribosomes are purified.
4. The mRNA fragments that are protected by the ribosomes are isolated and purified.
5. The purified mRNA fragments are sequenced using next-generation sequencing technology.
6. The resulting sequence data is analyzed to determine which regions of the mRNA are being actively translated.

Ribosome profiling has been used to study a variety of biological processes, including gene expression regulation, translational control, and viral infection. It has also been used to identify novel coding and non-coding RNA molecules.