Optimization of Ribosome Footprinting Conditions for Ribo-Seq in Human and

Autor: Katerina, Douka, Michaela, Agapiou, Isabel, Birds, Julie L, Aspden
Rok vydání: 2021
Zdroj: Frontiers in molecular biosciences. 8
ISSN: 2296-889X
Popis: Our understanding of mRNA translation and its regulation has been transformed by the development of ribosome profiling. This approach relies upon RNase footprinting of translating ribosomes in a precise manner to generate an accurate snapshot of ribosome positions with nucleotide resolution. Here we tested a variety of conditions, which contribute to the preciseness of ribosome footprinting and therefore the success of ribosome profiling. We found that NaCl concentration, RNaseI source, RNaseI amount, and temperature of footprinting all contributed to the quality of ribosome footprinting in human neuroblastoma SH-SY5Y cells. These ideal conditions for footprinting also improved footprint quality when used with
Databáze: OpenAIRE