期刊
GENOME BIOLOGY AND EVOLUTION
卷 10, 期 2, 页码 616-622出版社
OXFORD UNIV PRESS
DOI: 10.1093/gbe/evy022
关键词
GC content; GC enrichment; high-throughput sequencing; bird
资金
- Agence Nationale de la Recherche [ANR-14-CE02-0002-01]
- Agence Nationale de la Recherche (ANR) [ANR-14-CE02-0002] Funding Source: Agence Nationale de la Recherche (ANR)
Standard Illumina libraries are biased toward sequences of intermediate GC-content. This results in an underrepresentation of GC-rich regions in sequencing projects of genomes with heterogeneous base composition, such as mammals and birds. We developed a simple, cost-effective protocol to enrich sheared genomic DNA in its GC-rich fraction by subtracting AT-rich DNA. This was achieved by heating DNA up to 90 A degrees C before applying Illumina library preparation. We tested the new approach on chicken DNA and found that heated DNA increased average coverage in the GC-richest chromosomes by a factor up to six. Using a Taq polymerase supposedly appropriate for PCR amplification of GC-rich sequences had a much weaker effect. Our protocol should greatly facilitate sequencing and resequencing of the GC-richest regions of heterogeneous genomes, in combination with standard short-read and long-read technologies.
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