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Questions related from Gary Robertson
When I perform western blot experiments I often get inconsistent results post transfer. What I mean is that when I look at my PVDF membrane after the transfer step I can often see the molecular...
09 September 2018 4,896 4 View
To test if my double digest of the pTriex-3 vector is working I ran my 'uncut plasmid', 'double digested plasmid' and 'enzyme 1 cut plasmid' and 'enzyme 2 cut plasmid' on an agarose gel. Double...
03 March 2018 4,269 2 View
Short version: My plasmid contained insert but later after a second transformation into a second cell line the insert vanished and does not show up on PCR or sequencing Long version: I used...
02 February 2018 6,717 9 View
My BL21 cells when grown on an Ampicillin agar plate, completely cover the plate in a lawn as fast as 5 hours. They do this regardless of whether I transform them or not. A plate treated with no...
02 February 2018 367 5 View
I have a gene of interest approximately 2000 bp long coding for a protein approx. 670 amino acids in length. I wish to excise one of the proteins domains from the gene. To do so I need to remove...
09 September 2017 2,065 6 View
How long must a species last for, for it be detectable through typical absorbance spectroscopy. Say I have an enzyme with a turnover number of 300. Each reaction would take only 3.3 milli seconds....
08 August 2016 1,154 4 View
I am looking at an enzymes reaction mechanism. I have investigated 2 mutations. both mutations individually reduce the enzymes catalytic activity. The 2 mutations combined render any product...
04 April 2016 2,752 10 View
If I know that the free energy barrier of the first step of a enzymatic reaction has increased (And I know by how much) but the other steps remain unchanged may I infer from this any kind of...
01 January 2016 3,408 7 View
https://www.ncbi.nlm.nih.gov/pubmed/2742854I am following the method of Graminski in the paper above. Whether GSH is protonated or not may be followed with absorbance spectroscopy at 239 nm. To...
01 January 2016 3,790 4 View
I am doing an Nde1 and BamH1 double digest. The gel is as followed Pet-28a with no insert (undigested), pet-28a (digested),Thermoscientific zip ruler express 2, pet-11a with insert (undigested),...
07 July 2014 495 7 View
I am using GE healthcare poly-buffer 96 and PBE 94. I use a 0.025 M ethanolamine start buffer pH 9.4 and an elution buffer of PB 96 pH 6.0. I take care to degas all my buffers, keep the...
07 July 2014 2,731 2 View
I would like to compare two homo-dimers. Both Share one identical subunit (which I have aligned structurally) which binds to a second subunit. The specific piece of data I would like to compare is...
05 May 2014 5,734 6 View
Can someone please suggest a good programme that will either run on-line through a server on on windows that can calculate the ASA, planarity and Gap volume Index of a protein interface. I would...
05 May 2014 6,993 4 View
I need a way to create a simple to understand but informative diagram or schematic of a protein-protein interface. I am aware of the program Dimplot but would prefer to try something else. Any...
04 April 2014 5,861 3 View
