Facile, High Quality Sequencing of Bacterial Genomes from Small Amounts of DNA
Abstract
Sequencing bacterial genomes has traditionally required large amounts of genomic DNA (~1 μg). There have been few studies to determine the effects of the input DNA amount or library preparation method on the quality of sequencing data. Several new commercially available library preparation methods enable shotgun sequencing from as little as 1 ng of input DNA. In this study, we evaluated the NEBNext Ultra library preparation reagents for sequencing bacterial genomes. We have evaluated the utility of NEBNext Ultra for resequencing andde novoassembly of four bacterial genomes and compared its performance with the TruSeq library preparation kit. The NEBNext Ultra reagents enable high quality resequencing andde novoassembly of a variety of bacterial genomes when using 100 ng of input genomic DNA. For the two most challenging genomes (Burkholderiaspp.), which have the highest GC content and are the longest, we also show that the quality of both resequencing andde novoassembly is not decreased when only 10 ng of input genomic DNA is used.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Jan 01, 2014
- Source ID
- 10.1155/2014/434575
Entities
People
- Ayesha Arefin
- Beverly Parson-quintana
- Cheryl Gleasner
- Jennifer Price
- Karen W Davenport
- Kim Mcmurry
- Matthew Scholz
- Momchilo Vuyisich
- Patrick S Chain
- Shihai Feng
Organizations
- Defense Threat Reduction Agency
- Los Alamos National Laboratory
- Parkview Medical Center