I want to link two proteins with the linker (G4S)3. The linked (fused) proteins will then be expressed in E. coli. I will therefore be grateful if anybody could recommend a suitable DNA codon for this linker.
Just check the bacterial codon usage and select those with the most frequency in bacteria.
MICA and its receptor NKG2D were expressed as an inclusion bodies in E coli. It was therefore denatured and renatured employing dialysis method of refolding. Carrying out ELISA of MICA and it...
31 December 2013 7,562 7 View
I am trying to solubilize and refold an inclusion body of NKG2D with theoretical isoelectric point (PI) value of 6.2. I usually purify my solubilized protein with Nickel column (Elution buffer pH...
11 December 2013 9,778 5 View
I want to insert my recombinant DNA of about 1650bp into pET22b which is about 5500bp. The resultant product from the ligation will be used for E. coli transformation. I seem not to get a...
08 September 2013 9,957 12 View
NKG2D is a receptor expressed on NK and T cells. I would be grateful if anybody could suggest a suitable cell line that can express NKG2D on its surface for in vitro studies.
02 March 2013 7,998 0 View
I want to express these fused proteins and need to know which of the proteins should come first. This is to guide me in the primer design. MIC-A is a ligand of NKG2D and ScFv is a single chain...
31 December 2012 9,644 2 View
I am studying the bioactivy of two fused proteins. In order for the fused proteins to retain their respective bioactivities, I have decided to fuse them with a linker at their C-termini. I still...
31 December 2012 6,366 18 View
I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance
03 March 2021 3,568 1 View
I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...
01 March 2021 8,544 1 View
We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...
01 March 2021 3,355 3 View
I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....
01 March 2021 3,607 3 View
I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...
27 February 2021 547 3 View
Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...
26 February 2021 5,029 2 View
Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...
25 February 2021 6,969 3 View
I am trying to better understand the scope of DNA replication and sequencing errors, e.g. 1. I have seen similar error rates of 10e4 to 10e5 for cell & instrumental DNA replication,...
24 February 2021 4,397 3 View
I got a thin band and a thick band i guess it would be the genomic DNA and the 260/280 ratio is 3.
21 February 2021 6,523 6 View
I constructed a synthetic DNA library (scFv, VH-VL orientation) with a 3' reporter and 8x histag. I cloned and expressed this gene following which I performed an ELISA. The ELISA results suggested...
19 February 2021 7,006 5 View