A method to simultaneously construct up to 12 differently sized Illumina Nextera long mate pair libraries with reduced DNA input, time, and cost

Biotechniques. 2015 Jul 1;59(1):42-5. doi: 10.2144/000114310. eCollection 2015 Jul.

Abstract

Long mate pair (LMP) or "jump" libraries are invaluable for producing contiguous genome assemblies and assessing structural variation. However the consistent production of high quality (low duplication rate, accurately sized) LMP libraries has proven problematic in many genome projects. Input DNA length and quantity are key issues that can affect success. Here we demonstrate how 12 libraries covering a wide range of jump sizes can be constructed from <10 µg of DNA, thus ensuring production of the best LMP libraries from a given DNA sample. Finally, we demonstrate the accuracy of the insert sizes by mapping reads from each library back to an existing assembly.

Keywords: genome assembly; insert sizes; long mate pair (LMP) construction; wheat.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromosome Mapping / methods*
  • Costs and Cost Analysis
  • DNA, Plant / chemistry
  • Gene Library*
  • High-Throughput Nucleotide Sequencing
  • Nucleic Acid Amplification Techniques
  • Sequence Analysis, DNA
  • Time Factors
  • Triticum / genetics

Substances

  • DNA, Plant