ClickSeq: Replacing fragmentation and enzymatic ligation with click-chemistry to prevent sequence chimeras

Elizabeth Jaworski, Andrew Routh

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

We recently reported a fragmentation-free method for the synthesis of Next-Generation Sequencing libraries called “ClickSeq” that uses biorthogonal click-chemistry in place of enzymes for the ligation of sequencing adaptors. We found that this approach dramatically reduces artifactual chimera formation, allowing the study of rare recombination events that include viral replication intermediates and defective-interfering viral RNAs. ClickSeq illustrates how robust, bio-orthogonal chemistry can be harnessed in vitro to capture and dissect complex biological processes. Here, we describe an updated protocol for the synthesis of “ClickSeq” libraries.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages71-85
Number of pages15
Volume1712
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1712
ISSN (Print)1064-3745

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Keywords

  • Click-chemistry
  • ClickSeq
  • Flock house virus
  • Next-generation sequencing
  • RNAseq

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Jaworski, E., & Routh, A. (2018). ClickSeq: Replacing fragmentation and enzymatic ligation with click-chemistry to prevent sequence chimeras. In Methods in Molecular Biology (Vol. 1712, pp. 71-85). (Methods in Molecular Biology; Vol. 1712). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7514-3_6