Team:Cambridge/Protocols
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=Protocols= | =Protocols= | ||
- | + | ===Construct production=== | |
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- | + | * [[Team:Cambridge/Protocols/GelElectrophoresis|<u><span style="color:#00000CD">Gel Electrophoresis</span></u>]] A technique for separating DNA strands of different lengths. | |
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- | * [[Team:Cambridge/Protocols/GelElectrophoresis|<u><span style="color:#00000CD">Gel Electrophoresis</span></u>]] A technique for separating DNA strands of different lengths. | + | |
* [[Team:Cambridge/Protocols/GelExtractionofDNA|<u><span style="color:#00000CD">Gel Extraction of DNA</span></u>]] A technique used to isolate a desired fragment of intact DNA from an agarose gel following agarose gel electrophoresis. | * [[Team:Cambridge/Protocols/GelExtractionofDNA|<u><span style="color:#00000CD">Gel Extraction of DNA</span></u>]] A technique used to isolate a desired fragment of intact DNA from an agarose gel following agarose gel electrophoresis. | ||
* [[Team:Cambridge/Protocols/Gibsonassembly|<u><span style="color:#00000CD">Gibson Assembly</span></u>]] A technique for ligating multiple DNA fragments in one step, compatible with standard assembly. | * [[Team:Cambridge/Protocols/Gibsonassembly|<u><span style="color:#00000CD">Gibson Assembly</span></u>]] A technique for ligating multiple DNA fragments in one step, compatible with standard assembly. | ||
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* [[Team:Cambridge/Protocols/PCRProtocol|<u><span style="color:#00000CD">PCR using a high temperature DNA polymerase</span></u>]] A method for amplifying a section of DNA. | * [[Team:Cambridge/Protocols/PCRProtocol|<u><span style="color:#00000CD">PCR using a high temperature DNA polymerase</span></u>]] A method for amplifying a section of DNA. | ||
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+ | ===Transformation protocols=== | ||
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+ | * [[Team:Cambridge/Protocols/Chemicallycompetentcells|<u><span style="color:#00000CD">Chemically competent cells generation</span></u>]] A technique to produce e.coli cellsreceptive to chemical transformation. | ||
+ | * [[Team:Cambridge/Protocols/Electrocompetentcells|<u><span style="color:#00000CD">Electocompetent cells generation</span></u>]] A technique to produce e.coli cells receptive to transformation by electroporation | ||
+ | * [[Team:Cambridge/Protocols/ElectricalTransformation|<u><span style="color:#00000CD">Electroporation</span></u>]] A method for transforming appropriately competent cells with plasmid DNA using electricity. | ||
+ | * [[Team:Cambridge/Protocols/GlycerolStocks|<u><span style="color:#00000CD">Glycerol stocks</span></u>]] A technique for long term storage of cells at -80 degrees without losing cell vitality. | ||
* [[Team:Cambridge/Protocols/TransformationofB.subtilis|<u><span style="color:#00000CD">Transformation of ''Bacillus subtilis''</span></u>]] A technique used to introduce foreign DNA into competent Bacillus cells. | * [[Team:Cambridge/Protocols/TransformationofB.subtilis|<u><span style="color:#00000CD">Transformation of ''Bacillus subtilis''</span></u>]] A technique used to introduce foreign DNA into competent Bacillus cells. | ||
* [[Team:Cambridge/Protocols/TransformationofE.coli|<u><span style="color:#00000CD">Transformation of ''Escherichia coli''</span></u> ]] A method for transforming competent ''E.coli'' with DNA | * [[Team:Cambridge/Protocols/TransformationofE.coli|<u><span style="color:#00000CD">Transformation of ''Escherichia coli''</span></u> ]] A method for transforming competent ''E.coli'' with DNA | ||
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+ | ===Construct verification=== | ||
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+ | * [[Team:Cambridge/Protocols/PCRcolony|<u><span style="color:#00000CD">Colony PCR</span></u>]] PCR with cells as a template. Useful for checking the length of an insert in an introduced plasmid. | ||
+ | * [[Team:Cambridge/Protocols/RestrictionDigest|<u><span style="color:#00000CD">Restriction Enzyme Digest</span></u>]] A method for creating a restriction map of a plasmid. | ||
+ | * [[Team:Cambridge/Protocols/MiniPrep|<u><span style="color:#00000CD">MiniPrep - DNA extraction</span></u>]] A method used to extract DNA from bacterial cells. | ||
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+ | ===Ribosense testing=== | ||
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+ | * [[Team:Cambridge/Protocols/beta-galactosidaseassay|<u><span style="color:#00000CD">β-galactosidase assay</span></u>]] Assay to measure the amount of the enzyme β-galactosidase being produced by a population of cells. Useful as a reporter system. | ||
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+ | ===Ratiometrica testing=== | ||
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+ | * [[Team:Cambridge/Protocols/IPTGInduction|<u><span style="color:#00000CD">IPTG induction</span></u>]] A technique for inducing the pSPANK promoter in E.coli cells. | ||
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+ | ===Miscellaneous=== | ||
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+ | * [[Team:Cambridge/Protocols/biobrick_protocols|<u><span style="color:#00000CD">BioBricks</span></u>]] Resources for manipulation of BioBricks and information regarding the distribution. | ||
+ | * [[Team:Cambridge/Protocols/Plates|<u><span style="color:#00000CD">LB Agar Plates preparation</span></u>]] A method used to prepare agar plate to culture common bacteria. | ||
+ | * [[Team:Cambridge/Protocols/SDSPAGE|<u><span style="color:#00000CD">SDS PAGE protein analysis</span></u>]] A method used to separate polypeptides of different lengths. | ||
==Recipes== | ==Recipes== |
Revision as of 10:44, 3 September 2012
Contents |
Protocols
Construct production
- Gel Electrophoresis A technique for separating DNA strands of different lengths.
- Gel Extraction of DNA A technique used to isolate a desired fragment of intact DNA from an agarose gel following agarose gel electrophoresis.
- Gibson Assembly A technique for ligating multiple DNA fragments in one step, compatible with standard assembly.
- PCR using a high temperature DNA polymerase A method for amplifying a section of DNA.
Transformation protocols
- Chemically competent cells generation A technique to produce e.coli cellsreceptive to chemical transformation.
- Electocompetent cells generation A technique to produce e.coli cells receptive to transformation by electroporation
- Electroporation A method for transforming appropriately competent cells with plasmid DNA using electricity.
- Glycerol stocks A technique for long term storage of cells at -80 degrees without losing cell vitality.
- Transformation of Bacillus subtilis A technique used to introduce foreign DNA into competent Bacillus cells.
- Transformation of Escherichia coli A method for transforming competent E.coli with DNA
Construct verification
- Colony PCR PCR with cells as a template. Useful for checking the length of an insert in an introduced plasmid.
- Restriction Enzyme Digest A method for creating a restriction map of a plasmid.
- MiniPrep - DNA extraction A method used to extract DNA from bacterial cells.
Ribosense testing
- β-galactosidase assay Assay to measure the amount of the enzyme β-galactosidase being produced by a population of cells. Useful as a reporter system.
Ratiometrica testing
- IPTG induction A technique for inducing the pSPANK promoter in E.coli cells.
Miscellaneous
- BioBricks Resources for manipulation of BioBricks and information regarding the distribution.
- LB Agar Plates preparation A method used to prepare agar plate to culture common bacteria.
- SDS PAGE protein analysis A method used to separate polypeptides of different lengths.
Recipes
- CCMB80 for chemically competent cells generation A recipe for making CCMB80
- TAE buffer for gel electrophoresis A recipe for making 10xTAE buffer
- SOB growth medium A recipe for making SOB growth medium
Safety
See our Safety Page for Associated risk assessments for the above protocols and MSDS sheets for the reagents we have used.
Lab supplies
Other resources
File:Concentration calculator for lazy scientists.xls