alsA gene has two sub-units and some part of one sub-unit is bound to the TMD (alsC).
I am working on a thermophilic protein (pI: 7.72) purified using Ni-NTA chromatography (Final buffer: 25mM Pipes (6.6), 300mM NaCl, 1mM PMSF, 10% glycerol, 200mM imidazole). But the protein...
09 October 2016 8,338 24 View
I am working on a thermophilic protein which is not known to be toxic. It was earlier cloned in pET11a and cell death was observed after 2-3 hrs of IPTG (0.1mM) induction. I then cloned the gene...
06 July 2015 803 4 View
While eluting two of my proteins with 250mM of Imidazole, I am getting two different bands (Size of the proteins are mentioned in the Image attached). The genes are cloned in pET22b without Signal...
06 July 2015 1,772 8 View
In some papers people have recommended to use incubation at 42 degree celsius for short time for protein expression after IPTG induction? If YES, What will be its effect on protein's...
11 December 2014 3,484 5 View
I am trying to clone a T.thermophilus gene. I have used 3%/5% DMSO during PCR amplification (using phusion enzyme). Amplification is coming but after PCR purification, and I am also getting...
05 June 2014 2,234 21 View
I have tried cloning of my gene of interest in pET22b and pET28a, for different ratio, but the ligation didn't work (though colonies are coming in some cases). I am pretty much sure that my PCR...
04 May 2014 6,663 14 View
When I am doing PCR clean up after PCR amplification (using phusion + DMSO) of a thermophilus gene, and along with the amplified gene, the primer dimer is also appearing. Do you have any...
03 April 2014 1,451 18 View
I am trying to purify a protein from a thermophilic bacteria (gene cloned in DH5 alpha) but I am not getting expression of my protein after IPTG induction. I have tried with different expression...
02 March 2014 3,023 11 View
I am amplifying a gene from pET11a vector. Is the temperature for denaturation (95 C for chromosomal DNA generally) different for amplifying the gene from plasmid DNA?
02 March 2014 6,313 5 View
What are the other substances which can be used that give a positive result with alkaline picrate?
10 November 2013 8,908 3 View
I'm very interested in biotechnology and medical research and I'd like to meet a professional here and talk about it together
28 July 2024 3,798 4 View
List down the various factors responsible and how does this effect the overall biogas production when it is related to Hydraulic retention time of the digester based on the feedstock?
27 July 2024 4,800 4 View
I am pleased to announce the CUSABIO Biotechnology Scholarships for Students 2024! This scholarship aims to support outstanding students pursuing studies in the field of biotechnology. Here are...
22 July 2024 5,790 1 View
I was wondering if it is possible to form a permanent open "ssDNA bubble" similar to a transcription bubble (>13 nucleotides) within E. coli. These criteria are important: 1. Open ssDNA...
07 July 2024 8,275 5 View
Give in detail with available literature and website links for submissions
27 June 2024 9,753 1 View
Hey everyone! I'm having difficulty marking the C4 complement factor in Western Blotting. Has anyone had contact with this antibody or made it in-house? I have already used Anti-C4β (D-12)...
17 June 2024 6,124 1 View
Last week, I transformed home-prepared E. coli TOP10 cells. When checking the colonies, I noticed that some of them have yellow color. They did not grow on the randomly, but dependently on the...
16 June 2024 3,272 3 View
Hello, I have currently been tasked with performing a split GFP analysis. I have been using DH5a cells for plasmid transformation. The results were turning out to be what we expected. The...
29 May 2024 5,791 0 View
Generally for bacterial DNA isolation we use a fresh culture of 2-5 days old. But if we use an older culture say 10-15 days old will it have any impact on the DNA content and the DNA isolation...
13 May 2024 3,061 11 View
I am observing cleavage in my construct expressed in E. coli and attempting to identify the responsible proteases. Mass spectrometry analysis has revealed the sizes of the fragments, enabling...
12 May 2024 6,540 2 View