Methods: Difference between revisions
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Here you can find a list of methods for working with ''Bacillus subtilis'': | Here you can find a list of methods for working with ''Bacillus subtilis'': | ||
== Media == | |||
* [[C minimal medium]] | |||
* [[M9 minimal medium]] | |||
* [[Spizizen minimal medium]] | |||
== A collection of mutant strains == | |||
* the complete mutant strain collection from the [http://www.sciencedirect.com/science/article/pii/S2405471216304471?via%3Dihub Koo et al.] paper is now available: get access [https://www.addgene.org/kits/grosslab-bsubtilis-collections/ here] | |||
== Toolboxes for the work with ''B. subtilis'' == | |||
* the biobrick box {{PubMed|24295448}} | |||
* the biobrick box 2.0 {{PubMed|29118374}} | |||
* a part toolbox to tune genetic expression {{PubMed|27402159}} | |||
==Analysis of protein-protein interactions== | ==Analysis of protein-protein interactions== | ||
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* ''in vivo'' detection of [[protein-protein interactions]] using [[DivIVA]] and GFP {{PubMed|19698693}} | * ''in vivo'' detection of [[protein-protein interactions]] using [[DivIVA]] and GFP {{PubMed|19698693}} | ||
==Analysis of membrane proteins== | |||
* application of a split green fluorescent protein reporter {{PubMed|22285978}} | |||
== Antibodies == | |||
* List of available [[Antibodies]] | |||
== Protein expression == | |||
* the LIKE system {{PubMed|23110498}} | |||
==Strain construction== | ==Strain construction== | ||
* introduction of markerfree deletions: {{PubMed|20057169}} | * introduction of markerfree deletions: {{PubMed|26238998,24140578,23916947,23911571,22422839,20057169}} | ||
* generation of markerfree mutations: {{PubMed|20134232,21542786,24282588}} | |||
* generation of markerfree mutations: {{PubMed|20134232}} | * an improved method for transformation: {{PubMed|24838881}} | ||
==RNA analysis== | ==RNA analysis== | ||
* Mechanical cell disruption and extraction of RNA from ''B. subtilis'': [[Media:SOP-RNA.pdf]] | * 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|20300532}} | * transformation of wild-type ''B. subtilis'' strains incl. NCIB3610: {{PubMed|20300532}} | ||
* efficient electroporation of ''B. subtilis'' {{PubMed|22387342}} | |||
* a novel transformation protocol for ''B. subtilis'' DB104 {{PubMed|22395911}} | |||
==General procedures== | ==General procedures== | ||
* Standard operation procedures ([[SOP]]s) of the SYSMO-BACELL consortium | * Standard operation procedures ([[SOP]]s) of the SYSMO-BACELL consortium | ||
== | ==Microscopic techniques== | ||
* Live cell imaging of ''B. subtilis'' cells using automated time-lapse microscopy {{PubMed|21841760}} | * Live cell imaging of ''B. subtilis'' cells using automated time-lapse microscopy {{PubMed|21841760}} | ||
* Assay of gene expression dynamics using live cell imaging {{PubMed|22179594}} | * Assay of gene expression dynamics using live cell imaging {{PubMed|22179594}} | ||
* Visualization and quantification of gene expression heterogeneity in growing microbial cells {{PubMed|23874729}} | |||
* GFP variants specifically optimizd for use in ''B. subtilis'' {{PubMed|23956387}} | |||
* Quantitative analysis of cell types by comparing microscoy images {{PubMed|25448819}} | |||
* Semi-automated single cell analysis using membrane or DNA staining {{PubMed|27008090}} | |||
==Transposon mutagenesis== | ==Transposon mutagenesis== | ||
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==Protein-DNA interactions== | ==Protein-DNA interactions== | ||
* ChAP-chip: A modified ChIP-chip protocol for the ''in vivo'' identification of binding sites of DNA-binding proteins {{PubMed|17932079}} | * ChAP-chip: A modified ChIP-chip protocol for the ''in vivo'' identification of binding sites of DNA-binding proteins {{PubMed|17932079}} | ||
==Genome engineering== | |||
* generalized bacterial genome editing using mobile group II introns and Cre-lox {{PubMed|24002656}} | |||
* genome engineering using a synthetic gene circuit {{PubMed|25552415}} | |||
* Ordered Gene Assembly in Bacillus subtilis (OGAB) {{PubMed|25990947}} | |||
* construction of a super-competent strain {{PubMed|26732353}} | |||
* use of the CRISPR-Cas system {{PubMed|27260361,27255973,27238023,27342565}} | |||
* A review on genome engineering: | |||
<pubmed>23340847 </pubmed> | |||
== Visual descriptions of methods for ''B. subtilis'' in the ''Journal of Visual Experimentation'' == | |||
<pubmed> 24473333 24457605 24300024 24300445 22371091 20142799 21841760 </pubmed> | |||
==Key references== | ==Key references== | ||
<pubmed>22179594 20057169 20134232 21542786 20300532 19698693 17932079 21841760 17994626 23956387 22285978,22113911 24002656 24282588 26238998 25990947 26732353</pubmed> | |||
<pubmed>22179594 20057169 20134232 21542786 20300532 19698693 17932079 21841760 17994626 22285978</pubmed> | |||
Latest revision as of 10:31, 10 November 2017
Here you can find a list of methods for working with Bacillus subtilis:
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
- 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: 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:
<pubmed>23340847 </pubmed>
Visual descriptions of methods for B. subtilis in the Journal of Visual Experimentation
<pubmed> 24473333 24457605 24300024 24300445 22371091 20142799 21841760 </pubmed>
Key references
<pubmed>22179594 20057169 20134232 21542786 20300532 19698693 17932079 21841760 17994626 23956387 22285978,22113911 24002656 24282588 26238998 25990947 26732353</pubmed>