Difference between revisions of "Methods"

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(Microscopic techniques)
(Genome engineering)
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* Ordered Gene Assembly in Bacillus subtilis (OGAB) {{PubMed|25990947}}
 
* Ordered Gene Assembly in Bacillus subtilis (OGAB) {{PubMed|25990947}}
 
* construction of a super-competent strain {{PubMed|26732353}}     
 
* construction of a super-competent strain {{PubMed|26732353}}     
 +
* use of the CRISPR-Cas system {{PubMed|27260361,27255973,27238023}}   
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* A review on genome engineering:
 
* A review on genome engineering:
 
<pubmed>23340847 </pubmed>
 
<pubmed>23340847 </pubmed>

Revision as of 11:27, 8 June 2016

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

Media

Analysis of protein-protein interactions

  • Bacterial Two Hybrid: BACTH

Analysis of membrane proteins

  • application of a split green fluorescent protein reporter PubMed

Antibodies

Protein expression

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