Reducing ligation bias of small RNAs in libraries for next generation sequencing.

ABSTRACT:

Background

The use of nucleic acid-modifying enzymes has driven the rapid advancement in molecular biology. Understanding their function is important for modifying or improving their activity. However, functional analysis usually relies upon low-throughput experiments. Here we present a method for functional analysis of nucleic acid-modifying enzymes using next generation sequencing.

Findings

We demonstrate that sequencing data of libraries generated by RNA ligases can reveal novel secondary structure preferences of these enzymes, which are used in small RNA cloning and library preparation for NGS. Using this knowledge we demonstrate that the cloning bias in small RNA libraries is RNA ligase-dependent. We developed a high definition (HD) protocol that reduces the RNA ligase-

SUBMITTER: Sorefan K 

PROVIDER: S-EPMC3489589 | biostudies-literature | 2012 May

REPOSITORIES: biostudies-literature