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Questions related from Eleanor Martin
I want to use overlap extension PCR to insert a domain into my construct. However I have just realised that upstream of the region where I want to insert the domain there are 19 amino acids...
23 May 2017 5,236 4 View
New to crystallography...I want to compare ligand binding in a crystal structure to one generated using NMR. I have tried the pdbsum and the pisa servers but the hydrogen bonds that are identified...
28 December 2016 1,569 3 View
I am looking to create a stable cell line using Calu-3 cells. My initial plan is to co-transfect with pBABE-puro and select with puromycin. Does anyone have any similar experience with Calu-3? I...
17 August 2016 3,442 5 View
My POI is a human protein. However we when we had the DNA sequence synthesized we codon optimised for S.cerevisae (for protein expression). However, I now wish to do some interactome studies in a...
21 July 2016 1,548 3 View
I am new to ITC. I am currently using the technique to assess binding between a protein and a peptide to which it binds. The protein has two binding sites for the peptide, however site two is...
14 June 2016 9,565 4 View
I am working with a construct containing only the C-terminal region of my POI. It is 42 residues long, and contains no aromatic residues (but does contain a number of basic amino acids). Following...
16 November 2015 4,086 4 View
The gene I am working with is toxic to bacteria and frequently causes random deletions/insertions when cloning in E.coli. Therefore I have cloned my construct directly into yeast (my expression...
07 August 2015 624 9 View
I am trying to remove the GFP tag from our construct using the NEB Q5 SDM kit. My primers are designed to flank the region to be deleted and when I look at my PCR products on a gel (before the...
23 April 2015 7,415 5 View
