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De novo assembly of the Indian blue peacock (Pavo cristatus) genome using Oxford Nanopore technology and Illumina sequencing

BACKGROUND: The Indian peafowl (Pavo cristanus) is native to South Asia and is the national bird of India. Here we present a draft genome sequence of the male blue peacock using Illumina and Oxford Nanopore technology (ONT). RESULTS: ONT sequencing gave ∼2.3-fold sequencing coverage, whereas Illumin...

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Autores principales: Dhar, Ruby, Seethy, Ashikh, Pethusamy, Karthikeyan, Singh, Sunil, Rohil, Vishwajeet, Purkayastha, Kakali, Mukherjee, Indrani, Goswami, Sandeep, Singh, Rakesh, Raj, Ankita, Srivastava, Tryambak, Acharya, Sovon, Rajashekhar, Balaji, Karmakar, Subhradip
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6511069/
https://www.ncbi.nlm.nih.gov/pubmed/31077316
http://dx.doi.org/10.1093/gigascience/giz038
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author Dhar, Ruby
Seethy, Ashikh
Pethusamy, Karthikeyan
Singh, Sunil
Rohil, Vishwajeet
Purkayastha, Kakali
Mukherjee, Indrani
Goswami, Sandeep
Singh, Rakesh
Raj, Ankita
Srivastava, Tryambak
Acharya, Sovon
Rajashekhar, Balaji
Karmakar, Subhradip
author_facet Dhar, Ruby
Seethy, Ashikh
Pethusamy, Karthikeyan
Singh, Sunil
Rohil, Vishwajeet
Purkayastha, Kakali
Mukherjee, Indrani
Goswami, Sandeep
Singh, Rakesh
Raj, Ankita
Srivastava, Tryambak
Acharya, Sovon
Rajashekhar, Balaji
Karmakar, Subhradip
author_sort Dhar, Ruby
collection PubMed
description BACKGROUND: The Indian peafowl (Pavo cristanus) is native to South Asia and is the national bird of India. Here we present a draft genome sequence of the male blue peacock using Illumina and Oxford Nanopore technology (ONT). RESULTS: ONT sequencing gave ∼2.3-fold sequencing coverage, whereas Illumina generated 150–base pair paired-end sequence data at 284.6-fold coverage from 5 libraries. Subsequently, we generated a 0.915-gigabase pair de novo assembly of the peacock genome with a scaffold N50 of 0.23 megabase pairs (Mb). We predict that the peacock genome contains 23,153 protein-coding genes and 75.3 Mb (7.33%) of repetitive sequences. CONCLUSIONS: We report a high-quality assembly of the peacock genome using a hybrid approach of sequences generated by both Illumina and ONT. The long-read chemistry generated by ONT was useful for addressing challenges related to de novo assembly, particularly at regions containing repetitive sequences spanning longer than the read length, and which could not be resolved with only short-read–based assembly. Contig assembly of Illumina short reads gave an N50 of 1,639 bases, whereas with ONT, the N50 increased by >9-fold to 14,749 bases. The initial contig assembly based on Illumina sequencing reads alone gave 685,241 contigs. Further scaffolding on assembled contigs using both Illumina and ONT sequencing reads resulted in a final assembly of 15,025 super-scaffolds, with an N50 of ∼0.23 Mb. Ninety-five percent of proteins predicted by homology matched with those in a public repository, verifying the completeness of our assembly. Like other phylogenetic studies of avian conserved genes, we found P. cristatus to be most closely related to Gallus gallus, followed by Meleagris gallopavo and Anas platyrhynchos. Compared with the recently published peacock genome assembly, the current, superior, hybrid assembly has greater sequencing depth, fewer non-ATGC sequences, and fewer scaffolds.
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spelling pubmed-65110692019-05-15 De novo assembly of the Indian blue peacock (Pavo cristatus) genome using Oxford Nanopore technology and Illumina sequencing Dhar, Ruby Seethy, Ashikh Pethusamy, Karthikeyan Singh, Sunil Rohil, Vishwajeet Purkayastha, Kakali Mukherjee, Indrani Goswami, Sandeep Singh, Rakesh Raj, Ankita Srivastava, Tryambak Acharya, Sovon Rajashekhar, Balaji Karmakar, Subhradip Gigascience Data Note BACKGROUND: The Indian peafowl (Pavo cristanus) is native to South Asia and is the national bird of India. Here we present a draft genome sequence of the male blue peacock using Illumina and Oxford Nanopore technology (ONT). RESULTS: ONT sequencing gave ∼2.3-fold sequencing coverage, whereas Illumina generated 150–base pair paired-end sequence data at 284.6-fold coverage from 5 libraries. Subsequently, we generated a 0.915-gigabase pair de novo assembly of the peacock genome with a scaffold N50 of 0.23 megabase pairs (Mb). We predict that the peacock genome contains 23,153 protein-coding genes and 75.3 Mb (7.33%) of repetitive sequences. CONCLUSIONS: We report a high-quality assembly of the peacock genome using a hybrid approach of sequences generated by both Illumina and ONT. The long-read chemistry generated by ONT was useful for addressing challenges related to de novo assembly, particularly at regions containing repetitive sequences spanning longer than the read length, and which could not be resolved with only short-read–based assembly. Contig assembly of Illumina short reads gave an N50 of 1,639 bases, whereas with ONT, the N50 increased by >9-fold to 14,749 bases. The initial contig assembly based on Illumina sequencing reads alone gave 685,241 contigs. Further scaffolding on assembled contigs using both Illumina and ONT sequencing reads resulted in a final assembly of 15,025 super-scaffolds, with an N50 of ∼0.23 Mb. Ninety-five percent of proteins predicted by homology matched with those in a public repository, verifying the completeness of our assembly. Like other phylogenetic studies of avian conserved genes, we found P. cristatus to be most closely related to Gallus gallus, followed by Meleagris gallopavo and Anas platyrhynchos. Compared with the recently published peacock genome assembly, the current, superior, hybrid assembly has greater sequencing depth, fewer non-ATGC sequences, and fewer scaffolds. Oxford University Press 2019-05-11 /pmc/articles/PMC6511069/ /pubmed/31077316 http://dx.doi.org/10.1093/gigascience/giz038 Text en © The Author(s) 2019. Published by Oxford University Press. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Data Note
Dhar, Ruby
Seethy, Ashikh
Pethusamy, Karthikeyan
Singh, Sunil
Rohil, Vishwajeet
Purkayastha, Kakali
Mukherjee, Indrani
Goswami, Sandeep
Singh, Rakesh
Raj, Ankita
Srivastava, Tryambak
Acharya, Sovon
Rajashekhar, Balaji
Karmakar, Subhradip
De novo assembly of the Indian blue peacock (Pavo cristatus) genome using Oxford Nanopore technology and Illumina sequencing
title De novo assembly of the Indian blue peacock (Pavo cristatus) genome using Oxford Nanopore technology and Illumina sequencing
title_full De novo assembly of the Indian blue peacock (Pavo cristatus) genome using Oxford Nanopore technology and Illumina sequencing
title_fullStr De novo assembly of the Indian blue peacock (Pavo cristatus) genome using Oxford Nanopore technology and Illumina sequencing
title_full_unstemmed De novo assembly of the Indian blue peacock (Pavo cristatus) genome using Oxford Nanopore technology and Illumina sequencing
title_short De novo assembly of the Indian blue peacock (Pavo cristatus) genome using Oxford Nanopore technology and Illumina sequencing
title_sort de novo assembly of the indian blue peacock (pavo cristatus) genome using oxford nanopore technology and illumina sequencing
topic Data Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6511069/
https://www.ncbi.nlm.nih.gov/pubmed/31077316
http://dx.doi.org/10.1093/gigascience/giz038
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