Like ompc, OmpA which outer membrane protein in e coli can be used to tag and express a larger gene like 1000bp?
I want to clone a gene in Pgemt vector having BamH1 frw enzyme and Sal1 frw enzyme. where as in Pgemt vector BamH1 is considered to be a enzyme which "Restriction Enzymes That Do Not Cut the...
10 November 2014 1,911 3 View
Can we use BL21D3 competent cell which was purchased from NEB as a starter culture stock? What I mean is, I want to use the chemically competent cell which was bought from NEB as a glycerol stock...
03 April 2014 2,446 3 View
In our lab we mainly work on E.coli. For the past 2 months we are getting contamination in our LB tubes (a thread like think is growing). We have contamination only in LB tubes, so we cleaned all...
02 March 2014 5,823 17 View
I have got my gene of interested in polyacrilamide gel. I want to elute that band and used that for further cloning, so please help in providing a protocol for elute my DNA in polyacrilamide gel.
10 November 2013 4,921 9 View
How to calculate 5*10^7 cells from bacteria which is having a particular OD after Induction...
10 November 2013 588 4 View
How to run a polyacrylamide gel for DNA less than 100bp and what type of staining should be used? Alos, how to elude that gene of interest in that polyacrylamide gel?
09 October 2013 6,793 11 View
My gene of interest is 91bp. I cloned the PCR product in to Pgemt vector and digested with 2 enzymes. I am getting blue white colonies. While screening the white colonies, I am able to get the...
09 October 2013 6,907 9 View
Seeking to amplify 2 genes: one is 441bp and another one is 88bp. The problem is that the annealing temp of the primer is 72degree.and the GC content of both frw and rev primers are 59.6 and 47.5...
08 September 2013 3,410 9 View
I'm working on selecting antibodies against a recombinant protein that has a His-tag. My idea is to first bind the recombinant protein to a HisTRAP column and then use this column for an affinity...
07 August 2024 505 3 View
Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...
06 August 2024 5,373 2 View
Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...
06 August 2024 1,989 3 View
I ran a SDS-page of a bacterial lysate and I want to quantify protein concentration in a specific band. I was thinking of using a standards ladder or make some standards are different...
05 August 2024 9,805 3 View
I am having an issue with my gel image where my PCR product is not appearing very bright on the gel. When I perform gel extraction, the A260/280 purity value is very low. I used the Qiagen gel...
05 August 2024 9,798 3 View
I have protein-membrane simulations (PDB, PSF, DCD) and have noticed that water molecules near the protein are not visible in the simulations. How can I fix this issue? Is there a way to place the...
04 August 2024 1,200 2 View
Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC
01 August 2024 967 3 View
I have an RNA-seq data that I have analysed using Limma-voom and have extracted the gene IDs, log2FC and the p-values. At p value < 0.05, I have over 10,000 DEGs, however, when I run the GO...
31 July 2024 225 2 View
Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...
31 July 2024 9,892 4 View
I am currently working on a project that involves extracting cell membranes, for which we disrupt the cells using sonication. During the initial extraction process, we add protease inhibitors to...
30 July 2024 7,077 1 View