Inexpensive Multiplexed Library Preparation for Megabase-Sized Genomes
Harvard University · Center for Systems Biology · +1 more institution
Abstract
Whole-genome sequencing has become an indispensible tool of modern biology. However, the cost of sample preparation relative to the cost of sequencing remains high, especially for small genomes where the former is dominant. Here we present a protocol for rapid and inexpensive preparation of hundreds of multiplexed genomic libraries for Illumina sequencing. By carrying out the Nextera tagmentation reaction in small volumes, replacing costly reagents with cheaper equivalents, and omitting unnecessary steps, we achieve a cost of library preparation of $8 per sample, approximately 6 times cheaper than the standard Nextera XT protocol. Furthermore, our procedure takes less than 5 hours for 96 samples. Several…
Citation impact
- FWCI
- 20.33
- Percentile
- 100%
- References
- 16
Authors
6- MBMichael Baym
Harvard University, Center for Systems Biology
- SKSergey Kryazhimskiy
Harvard University, Center for Systems Biology
- TDTami D. Lieberman
Center for Systems Biology, Harvard University
- HCHattie Chung
Center for Systems Biology, Harvard University
- MMMichael M. DesaiCorresponding
Harvard University, Center for Systems Biology
Topics & keywords
- Amplicon
- Genome
- Biology
- DNA sequencing
- Computational biology
- Deep sequencing
- Amplicon sequencing
- Pyrosequencing
Funding
- NSNational Science FoundationAwards: R01-GM104239, 0903013, 1313638, PHY 1313638
- APAlfred P. Sloan FoundationAwards: GM104239, PHY 1313638
- JSJames S. McDonnell FoundationAwards: PHY 1313638, GM104239
- NINational Institutes of HealthAwards: R01-GM081617, PHY 1313638, GM104239, 281891, GM081617, R01-GM104239
- DODivision of Mathematical Sciences