Methods

Here you can find a list of methods for working with Bacillus subtilis:

Media

• C minimal medium
• M9 minimal medium

Toolboxes for the work with B. subtilis

• the biobrick box PubMed
• a part toolbox to tune genetic expression PubMed

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:

Kevin M Esvelt, Harris H Wang
Genome-scale engineering for systems and synthetic biology.
Mol Syst Biol: 2013, 9;641
[PubMed:23340847] [WorldCat.org] [DOI] (I p)