Difference between revisions of "Methods"

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(Strain construction)
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==Strain construction==
 
==Strain construction==
* introduction of markerfree deletions: {{PubMed|23916947,23911571,22422839,20057169}}
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* introduction of markerfree deletions: {{PubMed|24140578,23916947,23911571,22422839,20057169}}
  
* generation of markerfree mutations: {{PubMed|20134232}} and {{PubMed|21542786}}
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* generation of markerfree mutations: {{PubMed|20134232,21542786}}
  
 
==RNA analysis==
 
==RNA analysis==

Revision as of 18:06, 17 November 2013

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

Protein expression

Strain construction

  • introduction of markerfree deletions: PubMed
  • generation of markerfree mutations: 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

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
  • A review on genome engineering:

Key references