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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2018.1
- 수록면
- 281 - 288 (8page)
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The emergence and widespread use of high-throughput sequencing technologies have promoted metagenomic studies on environmental or animal samples. Library construction for metagenome sequencing and annotation of the produced sequence reads are important steps in such studies and influence the quality of metagenomic data. In this study, we collected some marine mollusk samples, such as Crassostrea hongkongensis, Chlamys farreri, and Ruditapes philippinarum, from coastal areas in South China. These samples were divided into two batches to compare two library construction methods for shellfish viral metagenome. Our analysis showed that reverse-transcribing RNA into cDNA and then amplifying it simultaneously with DNA by whole genome amplification (WGA) yielded a larger amount of DNA compared to using only WGA or WTA (whole transcriptome amplification). Moreover, higher quality libraries were obtained by agarose gel extraction rather than with AMPure bead size selection. However, the latter can also provide good results if combined with the adjustment of the filter parameters. This, together with its simplicity, makes it a viable alternative. Finally, we compared three annotation tools (BLAST, DIAMOND, and Taxonomer) and two reference databases (NCBI’s NR and Uniprot’s Uniref). Considering the limitations of computing resources and data transfer speed, we propose the use of DIAMOND with Uniref for annotating metagenomic short reads as its running speed can guarantee a good annotation rate. This study may serve as a useful reference for selecting methods for Shellfish viral metagenome library construction and read annotation.
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참고문헌 (30)
참고문헌 신청
Altschul SF / 1990 / Basic local alignment search tool / J Mol Biol 215:403~410
Banach BS / 2009 / Human airway epithelial cell culture to identify new respiratory viruses: coronavirus NL63 as a model / J Virol Methods 156:19~26
Buchfink B / 2015 / Fast and sensitive protein alignment using DIAMOND / Nat Methods 12:59~60
Carter NP / 1992 / Degenerate oligonucleotide-primed PCR: general amplification of target DNA by a single degenerate primer / Genomics 13:718~725
Djikeng A / 2008 / Viral genome sequencing by random priming methods / BMC Genom 9:5
Banach BS / 2009 / Human airway epithelial cell culture to identify new respiratory viruses: coronavirus NL63 as a model / J Virol Methods 156:19~26
Buchfink B / 2015 / Fast and sensitive protein alignment using DIAMOND / Nat Methods 12:59~60
Carter NP / 1992 / Degenerate oligonucleotide-primed PCR: general amplification of target DNA by a single degenerate primer / Genomics 13:718~725
Djikeng A / 2008 / Viral genome sequencing by random priming methods / BMC Genom 9:5
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