A step-by-step guide to assemble a reptilian genome

Asier Ullate-Agote, Yingguang Frank Chan, Athanasia C Tzika*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

1 Citation (Scopus)

Abstract

Multiple technologies and software are now available facilitating the de novo sequencing and assembly of any vertebrate genome. Yet the quality of most available sequenced genomes is substantially poorer than that of the golden standard in the field: the human genome. Here, we present a step-by-step protocol for the successful sequencing and assembly of a high-quality snake genome that can be applied to any other reptilian or avian species. We combine the great sequencing depth and accuracy of short reads with the use of different insert size libraries for extended scaffolding followed by optical mapping. We show that this procedure improved the corn snake scaffold N50 from 3.7 kbp to 1.4 Mbp, currently making it one of the snake genomes with the longest scaffolds. Short guidelines are also given on the extraction of long DNA molecules from reptilian blood and the necessary modifications in DNA extraction protocols. This chapter is accompanied by a website ( www.reptilomics.org/stepbystep.html ), where we provide links to the suggested software, examples of input and output files, and running parameters.

Original languageEnglish
Title of host publicationAvian and Reptilian Developmental Biology
EditorsG. Sheng
PublisherHumana Press
Pages47-67
Number of pages21
ISBN (Electronic)978-1-4939-7216-6
ISBN (Print)978-1-4939-7215-9
DOIs
Publication statusPublished - 2017
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1650
ISSN (Print)1064-3745

Keywords

  • Animals
  • Genome
  • High-Throughput Nucleotide Sequencing/methods
  • Male
  • Reptiles/classification
  • Sequence Analysis, DNA/methods
  • Snakes/genetics

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