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Questions related to Biochemical Techniques
Hello, I am trying to see flag tag protein (clonned in pcDNA plasmid) in U2OS cell lines using immunofluorescence assay. My control (pcDNA+GFP) shows high transfection efficiency (>80%)....
09 September 2014 3,770 7 View
My ESI- shows: 1151.5 (minor peak) / 864.8 (major peak) / 576.8 (medium peak).
09 September 2014 8,285 2 View
Hello, I want to know that how to check sensitivity of PCR ?
09 September 2014 262 8 View
I have a problem with amplification of a specific gene. After gene specific primer designing, I amplify my desired gene with PCR but don't get any answer. My enzyme is DreamTaq DNA polymerase. I...
09 September 2014 5,024 10 View
I have a question regarding ChIP-qPCR normalization. I am very familiar with qPCR and gene expression analyses but very new to chromatin immunoprecipitation qPCR. When performing gene expression,...
09 September 2014 6,438 4 View
Like the percentage of G4 which represents a certain sedimentation coefficient in the sedimentation profile?
09 September 2014 4,652 1 View
I have been expressing and lysing this construct for years, and today I got a thick gel instead of a nice supernatent. I recently started using Magic Media instead of IPTG induction, but this has...
09 September 2014 5,841 5 View
I want to study Iron starvation in cyanobacteria Nostoc punctiforme. But I did not want to use iron chelator in culture media, because they can make some difficulty.
09 September 2014 2,152 5 View
I am doing transfection for luciferase assay. How can I determine the amount of plasmid encoding gene of interest and plasmid encoding luciferase? Many thanks!
09 September 2014 8,981 2 View
We have been trying to extract "good quality of DNA" using different extraction methods like Column, Phenol Chloroform Isoamyl Alcohol and AFA methods and we found that AFA works better than...
09 September 2014 9,062 2 View
06 September 2014 6,295 1 View
I have to detect PPi levels in multiple cell (mammalian) lines.
08 August 2014 7,646 1 View
For example, our lab has been experiencing problems with dry Lithium Bromide taking on water mass when we work with it. If we make a highly concentrated solution of this salt as a stock solution,...
08 August 2014 7,740 4 View
What is the best pH for this reaction? Reaction mechanism of glutaraldehyde and cellulose.
16 May 2014 9,363 1 View
My thesis is about isolation of biosurfactant producing halophilic streptomyces, and now I have isolated some bacteria that look like streptomyces (chalky white colony ). I am trying to find out...
05 May 2014 1,582 2 View
05 May 2014 1,016 1 View
In order to construct unphosporylate kinase, I have replaced phopothreonine with threonine from active crystal structure computationally, as there is no inactive crystal coordinates for my kinase....
05 May 2014 5,961 0 View
I am having a tough time figuring out problems in my PCR where I am using a set of mouse IL2 primers and amplifying a segment. I am getting amplification with no template as well. I changed all...
05 May 2014 4,186 22 View
After staining and destaining my gel, my samples combined with each other and I can't see boundary between our samples
04 April 2014 1,160 12 View
I always use 1:32 dilutions of my standard stock for drawing the standard curve; once I got unexpected low efficiency (calculated by ABI 7500 software), and I was wondering whether the calculated...
04 April 2014 784 23 View
The same set of PCR conditions including the cDNA tube once gave nice clear bands, but two days later there were absolutely no bands at all (i have repeated this twice). This pattern has occurred...
04 April 2014 4,315 5 View
I'm doing SDS page electrophoresis. My gel does polymerize, but still a small aqueous layer remains at the surface.
04 April 2014 410 6 View
I am trying to quantify the TRAF6 protein using commercially available ELISA kits. However, the assay is unable to detect normal levels in patients. Standards work as per the guidelines. Any tips...
04 April 2014 3,691 1 View
I want to see the interaction of DNA polymerase and its associated proteins in vitro. Which method I should use?
04 April 2014 6,005 2 View