Experiments

Here you can find the protocols we used to conduct our experiments. Details of protocol adjustments can be found in our notebook.

Protocols for Construct Workflow

This are the protocols we used to create bacteria containing our riboswitch and reporter constructs.

  1. Primer resuspension
  2. IDT gene fragment resuspension
  3. Golden Gate Assembly
  4. Preparation of chemically competent E. coli
  5. Heat shock transformation
  6. Plasmid preperation (alkalic lysis)
  7. Plasmid preperation (ammonium acetat precipitation)
  8. Plasmid sequencing

Reporter assays

Assays we used for the expression of our riboswitch reporter constructs and measurement of results

  1. Reporter assays in liquid culture
  2. LacZ assay (ONPG-galactosidase assay)
  3. mScarlet-i3 / sfGFP assay
  4. NanoLuc assay

Media Recipes

Recipes we used for our media.

  1. SOB and SOC media
  2. TB medium