Menu
2021年06月01日

Low-input long-read sequencing for complete microbial genomes and metagenomic community analysis

Author(s): Heiner, Cheryl and Oh, Steve and Hall, Richard

Microbial genome sequencing can be done quickly, easily, and efficiently with the PacBio sequencing instruments, resulting in complete de novo assemblies. Alternative protocols have been developed to reduce the amount of purified DNA required for SMRT Sequencing, to broaden applicability to lower-abundance samples. If 50-100 ng of microbial DNA is available, a 10-20 kb SMRTbell library can be made. The resulting library can be loaded onto multiple SMRT Cells, yielding more than enough data for complete assembly of microbial genomes using the SMRT Portal assembly program HGAP, plus base modification analysis. The entire process can be done in less than 3 days by standard laboratory personnel. This approach is particularly important for analysis of metagenomic communities, in which genomic DNA is often limited. From these samples, full-length 16S amplicons can be generated, prepped with the standard SMRTbell library prep protocol, and sequenced. Alternatively, a 2 kb sheared library, made from a few ng of input DNA, can also be used to elucidate the microbial composition of a community, and may provide information about biochemical pathways present in the sample. In both these cases, 1-2 kb reads with >99.9% accuracy can be obtained from Circular Consensus Sequencing.

Organization: PacBio
Year: 2016

View Conference Poster

咨询专家

如果您有疑问、需要查看订单状态或想要购买仪器,我们随时乐意提供帮助。

姓名(Required)
这个字段是用于验证目的,应该保持不变。

在本网页上注册,即表示您同意,并同意 PacBio 根据我们的隐私政策收集和使用该信息.