Determining degradation intermediates and the pathway of 3' to 5' degradation of histone mRNA using high-throughput sequencing.

ABSTRACT: The half-life of an mRNA is an important parameter contributing to the steady-state level of the mRNA. Rapid changes in mRNA levels can result from decreasing the half-life of an mRNA. Establishing the detailed pathway of mRNA degradation for a particular class of mRNAs requires the ability to isolate mRNA degradation intermediates. High-throughput sequencing provides a method for detecting these intermediates. Here we describe a method for determining the intermediates in 3' to 5' degradation. Characterizing these intermediates requires not only determining the precise 3' end of the molecule to a single nucleotide resolution, but also the ability to detect and characterize any untemplated nucleotides present on the intermediates. We achieve this by ligating a known sequence to all the 3'

SUBMITTER: Holmquist CE 

PROVIDER: S-EPMC7252413 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature