Methods
Here you can find a list of methods for working with Bacillus subtilis:
Contents
- 1 Media
- 2 A collection of mutant strains
- 3 Toolboxes for the work with B. subtilis
- 4 Analysis of protein-protein interactions
- 5 Analysis of membrane proteins
- 6 Antibodies
- 7 Protein expression
- 8 Strain construction
- 9 RNA analysis
- 10 Genetic work with non-standard strains
- 11 General procedures
- 12 Microscopic techniques
- 13 Transposon mutagenesis
- 14 Protein-DNA interactions
- 15 Genome engineering
- 16 Visual descriptions of methods for B. subtilis in the Journal of Visual Experimentation
- 17 Key references
Media
A collection of mutant strains
- the complete mutant strain collection from the Koo et al. paper is now available: get access here
Toolboxes for the work with B. subtilis
Analysis of protein-protein interactions
- Bacterial Two Hybrid: BACTH
- SPINE: A method to detect in vivo protein-protein interactions
- in vivo detection of protein-protein interactions using DivIVA and GFP PubMed
Analysis of membrane proteins
- application of a split green fluorescent protein reporter PubMed
Antibodies
- List of available Antibodies
Protein expression
- the LIKE system PubMed
Strain construction
- introduction of markerfree deletions: PubMed
- generation of markerfree mutations: PubMed
- an improved method for transformation: PubMed
RNA analysis
- Mechanical cell disruption and extraction of RNA from B. subtilis: Media:SOP-RNA.pdf
Genetic work with non-standard strains
- transformation of wild-type B. subtilis strains incl. NCIB3610: PubMed
- efficient electroporation of B. subtilis PubMed
- a novel transformation protocol for B. subtilis DB104 PubMed
General procedures
- Standard operation procedures (SOPs) of the SYSMO-BACELL consortium
Microscopic techniques
- Live cell imaging of B. subtilis cells using automated time-lapse microscopy PubMed
- Assay of gene expression dynamics using live cell imaging PubMed
- Visualization and quantification of gene expression heterogeneity in growing microbial cells PubMed
- GFP variants specifically optimizd for use in B. subtilis PubMed
- Quantitative analysis of cell types by comparing microscoy images PubMed
- Semi-automated single cell analysis using membrane or DNA staining PubMed
Transposon mutagenesis
- mariner transposon mutagenesis for random inducible-expression insertions and transcriptional reporter fusion insertions PubMed
Protein-DNA interactions
- ChAP-chip: A modified ChIP-chip protocol for the in vivo identification of binding sites of DNA-binding proteins PubMed
Genome engineering
- generalized bacterial genome editing using mobile group II introns and Cre-lox PubMed
- genome engineering using a synthetic gene circuit PubMed
- Ordered Gene Assembly in Bacillus subtilis (OGAB) PubMed
- construction of a super-competent strain PubMed
- use of the CRISPR-Cas system PubMed
- A review on genome engineering:
Visual descriptions of methods for B. subtilis in the Journal of Visual Experimentation
Key references
