Rapid Quantification of Mutant Fitness in Diverse Bacteria by Sequencing Randomly Bar-Coded Transposons
Lawrence Berkeley National Laboratory · Joint Genome Institute · +1 more institution
Abstract
UNLABELLED: Transposon mutagenesis with next-generation sequencing (TnSeq) is a powerful approach to annotate gene function in bacteria, but existing protocols for TnSeq require laborious preparation of every sample before sequencing. Thus, the existing protocols are not amenable to the throughput necessary to identify phenotypes and functions for the majority of genes in diverse bacteria. Here, we present a method, random bar code transposon-site sequencing (RB-TnSeq), which increases the throughput of mutant fitness profiling by incorporating random DNA bar codes into Tn5 and mariner transposons and by using bar code sequencing (BarSeq) to assay mutant fitness. RB-TnSeq can be used with any transposon, and…
Citation impact
- FWCI
- 23.43
- Percentile
- 100%
- References
- 39
Authors
11- KMKelly M. WetmoreCorresponding
Lawrence Berkeley National Laboratory
- MNMorgan N. Price
Lawrence Berkeley National Laboratory
- RJR. Jordan Waters
Lawrence Berkeley National Laboratory, Joint Genome Institute
- JSJacob S. Lamson
Lawrence Berkeley National Laboratory
- JHJennifer He
Lawrence Berkeley National Laboratory
Topics & keywords
- Transposon mutagenesis
- Biology
- Genetics
- Transposable element
- Computational biology
- Mutant
- Sleeping Beauty transposon system
- DNA sequencing
Funding
- UDU.S. Department of EnergyAwards: -AC02-05CH11231, 05CH11231, AC02-05CH11231, DE-AC02, DE-AC02-05CH11231, DE-AC02-
- JGJoint Genome InstituteAwards: DE-AC02-05CH11231, AC02-05CH11231
- OOOffice of ScienceAwards: AC02-05CH11231, -AC02-05CH11231, DE-AC02
- BABiological and Environmental ResearchAwards: 05CH11231, DE-AC02-05CH11231, AC02-05CH11231
- LDLaboratory Directed Research and DevelopmentAward: DE-AC02-05CH11231
- LBLawrence Berkeley National LaboratoryAwards: DE-AC02-05CH11231, 05CH11231, AC02-05CH11231