Related Experiment Video
Updated: Mar 30, 2026

Sequencing of mRNA from Whole Blood using Nanopore Sequencing
Published on: June 3, 2019
Identification of DNA lesions using a third base pair for amplification and nanopore sequencing
Jan Riedl1, Yun Ding1, Aaron M Fleming1
1Department of Chemistry, University of Utah, 315S 1400 East, Salt Lake City, Utah 84112-0850, USA.
Abstract:
Damage to the genome is implicated in the progression of cancer and stress-induced diseases. DNA lesions exist in low levels, and cannot be amplified by standard PCR because they are frequently strong blocks to polymerases. Here, we describe a method for PCR amplification of lesion-containing DNA in which the site and identity could be marked, copied and sequenced. Critical for this method is installation of either the dNaM or d5SICS nucleotides at the lesion site after processing via the base excision repair process. These marker nucleotides constitute an unnatural base pair, allowing large quantities of marked DNA to be made by PCR amplification. Sanger sequencing confirms the potential for this method to locate lesions by marking, amplifying and sequencing a lesion in the KRAS gene. Detection using the α-hemolysin nanopore is also developed to analyse the markers in individual DNA strands with the potential to identify multiple lesions per strand.
Related Concept Videos
Sanger Sequencing
Next-generation Sequencing
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
Maxam-Gilbert Sequencing
Challenges of the Maxam-Gilbert Method
The...
Long-patch Base Excision Repair

