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Questions related from Michael Swanson
We have an 1100 series HPLC with a quaternary pump that is an upgraded isocratic pump (G1310A) that will not power on. Everything else in the system powers on fine and I have tried moving the...
09 September 2017 6,446 8 View
I am using pET24a and I would like to use the NheI and XhoI sites to add our gene. If I use the NheI site at the N-terminal, the first methionine will be from the vector itself, and not the gene...
11 November 2016 7,952 7 View
We have our mitochondrial protein of interest in a mammalian expression vector with a C terminal Turbo GFP (from origene). We want to over express this protein in bacteria, purify it then add it...
05 May 2016 2,790 1 View
I'm trying to clone a ~500 bp gene into the pET28a plasmid. I first cloned this gene into the pCMV6-AC-DDK plasmid (using AscI and XhoI) to incorporate the DDK tag. This was successful verified...
04 April 2016 5,919 6 View
I have been trying to express a mammalian mitochondrial protein that is ~100kD. The gene is in the pET47b vector and I have been using the Rosetta2 (DE3) strain. I have done SDS-PAGE/westerns to...
07 July 2015 356 15 View
Does anyone know if a streptavidin agarose column will have any affinity for lipoic acid or more specifically, lipoylated proteins? We have a protein that is lipoylated for an assay and being...
01 January 2015 4,622 4 View
I am working on cloning a 3 kb gene into pET-47b using engineered restriction sites. Sequencing showed that my gene was successfully ligated into pET47 but it contains 7 missense mutations. I...
06 June 2014 6,458 10 View
