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Questions related from Martin Sim
I have 2 cell groups; (WT)Wild-Type (Subject)Wild-Type+Transfected Gene I expose these cells to a particular drug at different doses. I then look at calcium influx. The drug is known (has...
05 May 2019 2,414 4 View
I have 3 cells lines; Wild type control Wild type transfected with GFP vector Wild type transfected with GOI-GFPtag I'd like to do some general characterization on the cells, including cell...
06 June 2018 9,977 0 View
How vital is it to optimize the RNA input for reverse transcription for subsequent qPCR reactions? The qPCR reaction mix protocol calls for 25ng of template. I've been using 100ng total RNA for...
03 March 2018 641 5 View
I have plasmid containing my gene of interest. It has been linearized, gel purified, and is at known concentration. I will use this standard to generate a standard curve for my qPCR runs, and...
03 March 2018 9,203 4 View
I'm very familiar with neuroblastoma cell lines, both general culturing, and differentiating. However, I've been considering transfecting my gene of interest into a non-cancer cell line. What are...
03 March 2018 5,965 2 View
I'm considering performing a PCR on my gene sets (both GOIs and house keeping genes), run the PCR products on a gel, extract them from the gel, and then measure their concentrations (using a...
03 March 2018 9,726 5 View
I have RNA samples that I will be using for cDNA synthesis, and analysis of expression levels. The RNA has been DNAse treated, both in column during extraction, and post column extraction...
03 March 2018 826 1 View
I have acquired a lot of images of my neuron's axons degenerating over time by various different insults. A lot of my data is qualitative, and I could perhaps go through all the images and count...
02 February 2018 2,457 2 View
I grow my neurons on glass surface. Their axons so delicate that I cannot remove the media entirely. When I refeed them, I remove half the media and replace it with fresh media containing double...
02 February 2018 1,854 3 View
I've added 6-OHDA to my neuronal cultures to test whether they are dopaminergic, and they do die. However, I was told that I need to also inhibit noradrenaline reuptake trasnporters as 6-OHDA is...
01 January 2018 9,956 2 View
I performed an experiment involving two different treatments, and used both a negative and positive control. Every sample, including the controls was done in replicates. The experiment was...
11 November 2017 6,867 2 View
I grow my cells on both plastic Corning 96well culture plates for most experiments, but for immunoluorescence staining I'm growing them on glass or permanox chamber slides. One of the Chamber...
11 November 2017 4,752 4 View
I am looking for ordinary 96 well tissue culture plates, but with glass bottom. Please help.
11 November 2017 7,887 0 View
It's my first experiment in flow cytometry. I'm attempting to exclude non-single cells. I'm wondering why it's not common practice to simply discriminate using pulse width (Time of Flight)? What...
11 November 2017 9,151 4 View
As an example, the pEF vector available from Sigma, as well as most EF1 vectors on addgene have a 1172 bp EF1a promoter sequence. The SystemBio pCDH vectors have a smaller 545bp EF1a promoter...
06 June 2017 3,691 8 View
The EF1 pCDH vectors by SystemBio use a composite promoter; hEF1a-HTLV http://discovery.ucl.ac.uk/20480/1/20480.pdf "hEF1a-HTLV promoter is a composite promoter comprising the human Elongation...
06 June 2017 9,966 1 View
