Abstract
Single-molecule real-time (SMRT) sequencing developed by Pacific BioSciences (PacBio) offers three major advantages compared to second-generation sequencing: long read length and high consensus accuracy, and a low degree of bias. Together with high sequencing coverage, these advantages overcome the difficulty of sequencing genomic regions such as long AT-rich islands and repeated regions (e.g., ribosomal DNA) in the genome of Trichoderma reesei QM6a. Herein, we describe a protocol for preparing high-quality, high molecular weight genomic DNA for PacBio long-read sequencing, de novo assembly and streamlined annotation of the QM6a genome.
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Acknowledgments
We thank Shu-Yun Tung (IMB Genomic Core) for NGS sequencing service, John O′Brien for English editing, and Yu-Tang Huang (IMB Computer Room) for maintaining the computer workstation. We are grateful to Roland Martzy for his efforts in editing this book chapter. Funding from Academia Sinica, Taiwan, Republic of China [AS-105-TP-B07 and AS108-TP-B07] to TFW.
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Li, WC., Wang, TF. (2021). PacBio Long-Read Sequencing, Assembly, and Funannotate Reannotation of the Complete Genome of Trichoderma reesei QM6a. In: Mach-Aigner, A.R., Martzy, R. (eds) Trichoderma reesei. Methods in Molecular Biology, vol 2234. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1048-0_21
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DOI: https://doi.org/10.1007/978-1-0716-1048-0_21
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