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 qualitatively measure the amount of the enzyme β-galactosidase being produced by a population of cells. Useful as a reporter system.
• Miller assay Assay to quantify the amount of the enzyme β-galactosidase being produced by a population of cells. Useful as a quantitative reporter system.
• Production of magnesium free cells. Technique to produce cells that contain little or no magnesium, to control for this variable in tests involving the magnesium riboswitch.

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.
• Imaging sporesA method for seeing vegetative cells and bacillus spores under the microscope.

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
• CDSM A Chemically Defined Media to induce sporulation in B. Subtilis
• Germination Media A Chemically Defined Media to induce Germination in B. Subtilis

Safety

See our Safety Page for Associated risk assessments for the above protocols and MSDS sheets for the reagents we have used.

Lab supplies

• BioPath Stores
• Plant Department Stores Catalogue xls (Updated 2nd July 2012)

Other resources

File:Concentration calculator for lazy scientists.xls