Plasmid stability
I want to purify IgG4 antibodies from immunized rabbits. I have tried to look for protocols to no avail. Commercial kits on the other hand exist for humans and a few other animals but I have not...
03 April 2019 10,153 3 View
Hello everyone, we are characterizing some proteins in the context of vaccine development and we are interested in determining there are any epitopes that may rather lead to immune evasion. Don't...
03 April 2019 3,080 3 View
Hello everyone, so I expressed this protein and I am trying to further purify using SEC and still having the impurities in the fraction I expected to have the protein. How do I eliminate these. I...
03 April 2019 7,714 3 View
I am doing RNA, DNA and protein extraction from the same sample and my protocol recommends this water saturated phenol:chloroform mixture. I am not not very versed with how to go about with the...
03 April 2018 4,443 8 View
Hello everyone, I am trying to do some hands-on bioinformatics and will be grateful to get your orientation.
31 December 2017 1,479 5 View
Hello everyone, I did 3D structure prediction (using I-TASSER) for a protein and submitted the best model for structure refinement (with GalaxyRefine). Unfortunately, I don't have a very good...
11 December 2017 1,362 0 View
Hello everyone, I am about doing an antigen capture ELISA and a little confused about which buffer can be used in absorbing the capture antibody to the microplates. I am using Nunc MaxiSorb 96...
10 November 2017 2,991 3 View
Hello everyone, So I planned this ELISA to do, unfortunately I coated plates and for some reason I am not able to continue with the experiment as planned. So basically, instead of coating...
09 October 2017 9,378 4 View
Hello everyone, I wish to express a gene cloned in pGEX5x3 through auto-induction and compare with the normal IPTG induction. Any one has experience doing autoinduction with pGEX vectors?
08 September 2017 6,429 0 View
I am working with a pET30a+ construct and my protein is C-terminal His-tagged. My protein is forming inclusion bodies and I need to get a protocol for isolation of a pure protein from this. An...
07 August 2017 2,097 4 View
I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?
11 August 2024 5,138 1 View
I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...
08 August 2024 4,645 2 View
After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...
08 August 2024 1,668 3 View
I'm trying to find a DNA extraction method for fungi that does not require equipment and heating. Is there anyone who can suggest an alternative option? Thank you
08 August 2024 4,733 2 View
I have been working on Red blood cell-derived extracellular vesicles as Antisense Oligonucleotide (ASO) carriers. We normally run agarose gel to quantify the loading efficiency. I used naked ASO...
06 August 2024 3,130 2 View
Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...
05 August 2024 1,573 4 View
I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...
05 August 2024 9,059 3 View
I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....
05 August 2024 2,570 3 View
I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...
01 August 2024 2,079 0 View
Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC
01 August 2024 967 3 View
