Analysis of RNA base modification and structural rearrangement by single-molecule real-time detection of reverse transcription.

ABSTRACT:

Background

Zero-mode waveguides (ZMWs) are photonic nanostructures that create highly confined optical observation volumes, thereby allowing single-molecule-resolved biophysical studies at relatively high concentrations of fluorescent molecules. This principle has been successfully applied in single-molecule, real-time (SMRT®) DNA sequencing for the detection of DNA sequences and DNA base modifications. In contrast, RNA sequencing methods cannot provide sequence and RNA base modifications concurrently as they rely on complementary DNA (cDNA) synthesis by reverse transcription followed by sequencing of cDNA. Thus, information on RNA modifications is lost during the process of cDNA synthesis.

Results

Here we describe an application of SMRT technology to follow the activity of

SUBMITTER: Vilfan ID 

PROVIDER: S-EPMC3623877 | biostudies-literature | 2013 Apr

REPOSITORIES: biostudies-literature