Center for Phage Technology, Texas A&M University, College Station, TX, USA.
Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX, USA.
Methods Mol Biol. 2024;2738:125-144. doi: 10.1007/978-1-0716-3549-0_8.
Bacteriophages, or more simply phages, are currently experiencing a renaissance in life science research for their roles in natural microbial communities, their potential use as antimicrobials, and biotechnological applications. In the modern era, one of the primary steps in phage characterization is obtaining the sequence of the complete genome; this information can be used to determine the relationship of the phage to known phages, predict phage lifestyle, and is a prerequisite for many downstream applications. This protocol describes methods for determining the complete sequence of a double-stranded DNA bacteriophage genome, including DNA extraction from a phage lysate, sending the DNA out to a sequencing service, assembly of the sequence raw reads, and completion of the genome sequence.
噬菌体，或者更简单地称为 phage，由于其在自然微生物群落中的作用、作为抗菌剂的潜在用途以及生物技术应用，目前在生命科学研究中重新焕发生机。在现代，噬菌体表征的主要步骤之一是获得完整基因组的序列；该信息可用于确定噬菌体与已知噬菌体的关系、预测噬菌体的生活方式，并且是许多下游应用的前提。本方案描述了确定双链 DNA 噬菌体基因组完整序列的方法，包括从噬菌体裂解物中提取 DNA、将 DNA 发送到测序服务、组装序列原始读数以及完成基因组序列。
