Cloning and mutagenesis of BrxR protein
Corresponding Organization : Durham University
Other organizations : Roslin Institute, University of Edinburgh, New England Biolabs (United States), University of Liverpool
Variable analysis
- Primers TRB878 and TRB879 used to amplify brxR from pEFER
- Primers TRB876 and TRB877 used to amplify brxR from pEFER
- Primers TRB1987 and TRB1988 used to perform QuikChange mutagenesis to produce pBAD30-his6-brxR-R17A
- Production of pTRB446 via Ligation Independent Cloning (LIC)
- Production of pBAD30-his6-brxR
- Production of pBAD30-his6-brxR-R17A
- Monarch DNA kits (NEB) used for purification of PCR amplicons and plasmids
- Standard molecular biology techniques used for PCR, restriction digests, ligations, transformations and agarose gel electrophoresis
- Constructed plasmids confirmed via sequencing with an Abi 3370 DNA sequencer
- No positive or negative controls explicitly mentioned
Annotations
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