Methods

Media

• [wiki|C minimal medium]
• [wiki|M9 minimal medium]
• [wiki|Spizizen minimal medium]

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

• the biobrick box PubMed
• the biobrick box 2.0 PubMed
• a part toolbox to tune genetic expression PubMed

Analysis of protein-protein interactions

• Bacterial Two Hybrid: [wiki|BATCH]
• [wiki|SPINE]: A method to detect in vivo [wiki|Protein-protein interactions|protein-protein interactions]
• in vivo detection of protein-protein interaction using [protein|search|DivIA] and GFP  PubMed

Analysis of membrane proteins

application of a split green fluorescent protein reporter PubMed

Antibodies

List of available [wiki|Antibodies|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

Genetik 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 ([wiki|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

Genomic 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: PubMed

Visual descriptions of methods of B. subtilis in the Journal of Visual Experimentation

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Key references

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