Rxivist logo

Rxivist combines preprints from bioRxiv with data from Twitter to help you find the papers being discussed in your field. Currently indexing 70,653 bioRxiv papers from 308,421 authors.

Detection of Base Analogs Incorporated During DNA Replication by Nanopore Sequencing

By Daniela Georgieva, Qian Liu, Kai Wang, Dieter Egli

Posted 14 Feb 2019
bioRxiv DOI: 10.1101/549220

DNA synthesis is a fundamental requirement for cell proliferation and DNA repair, but no tools exist to identify the location, direction and speed of replication forks with base pair resolution. Mammalian cells have the ability to incorporate thymidine analogs along with the natural A, T, G and C bases during DNA synthesis, which allows for labelling of replicating or repaired DNA. The Oxford Nanopore Technologies (ONT) MinION infers nucleotide identity from changes in the ionic current as DNA strands are pulled through nanopores and can differentiate noncanonical nucleotides from natural ones. Here, we demonstrate the use of the ONT MinION to detect 11 different thymidine analogs including CldU, BrdU, IdU, as well as, EdU alone or coupled to Biotin and other bulky adducts in synthetic DNA templates. We also show detection of IdU label, incorporated during DNA replication in the genome of mouse pluripotent stem cells. We find that different modifications generate variable shifts in signals, providing a method of using analog combinations to identify the location and direction of DNA synthesis and repair at high resolution. We conclude that this novel method has the potential for single-base, genome-wide examination of DNA replication in stem cell differentiation or cell transformation.

Download data

  • Downloaded 994 times
  • Download rankings, all-time:
    • Site-wide: 8,345 out of 70,658
    • In genomics: 1,297 out of 4,718
  • Year to date:
    • Site-wide: 2,528 out of 70,658
  • Since beginning of last month:
    • Site-wide: 8,050 out of 70,658

Altmetric data


Downloads over time

Distribution of downloads per paper, site-wide


PanLingua

Sign up for the Rxivist weekly newsletter! (Click here for more details.)


News