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Whole genome sequencing of Theileria parva using target capture
Affiliation:1. Agricultural Research Council, Onderstepoort Veterinary Research, Private Bag X05, Onderstepoort, 0110 Pretoria, South Africa;2. Department of Veterinary Tropical Diseases, Vector and Vector-borne Disease Research Programme, University of Pretoria, Private Bag X04, Onderstepoort, 0110 Pretoria, South Africa;3. Agricultural Research Council, Biotechnology Platform, Private Bag X05, Onderstepoort, 0110 Pretoria, South Africa;4. School of Life Sciences, University of KwaZulu-Natal, Private Bag X54001, Durban 4000, South Africa;5. Department of Life and Consumer Sciences, University of South Africa, Florida 1709, South Africa
Abstract:
Protozoan parasite isolation and purification are laborious and time-consuming processes required for high quality genomic DNA used in whole genome sequencing. The objective of this study was to capture whole Theileria parva genomes directly from cell cultures and blood samples using RNA baits. Cell culture material was bait captured or sequenced directly, while blood samples were all captured. Baits had variable success in capturing T. parva genomes from blood samples but were successful in cell cultures. Genome mapping uncovered extensive host contamination in blood samples compared to cell cultures. Captured cell cultures had over 81 fold coverage for the reference genome compared to 0–33 fold for blood samples. Results indicate that baits are specific to T. parva, are a good alternative to conventional methods and thus ideal for genomic studies. This study also reports the first whole genome sequencing of South African T. parva.
Keywords:
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