19 Questions 14 Answers 0 Followers
Questions related from Frank Liu
After one harvests a mouse spleen and isolates the NK cells (through a commercial kit, for example) roughly how many NK cells can one expect to be left with? If anyone has rough numbers they can...
29 January 2022 5,455 2 View
My goal is to establish an in-vitro culture of mouse splenocytes that are lacking in either CD4 T cells or CD8 T cells. I tried doing this in the past by administering IP injections of anti-CD4...
14 January 2022 5,665 3 View
What sort of experiments would you conduct, or databases would you search, in order to identify all of the transcriptional targets of a certain transcription factor of interest? For instance, say...
10 June 2019 5,864 3 View
I am trying to monitor the methylation statuses of a certain set of genetic loci in glioblastoma cells following a treatment protocol. Can anyone please suggest some easy-to-use assay kits to...
14 March 2019 9,698 2 View
Is there any notable advantage/disadvantage or reason to use either propidium iodide, zombie red, or annexin V for flow cytometry detection of cell viability? Thanks.
02 June 2018 3,151 3 View
Hello all, I recently treated telomerase-inhibited A549 cells with 5 uM RITA, a p53 activator drug, and measured cell viability using the MTT assay. While wild type A549 experienced an expected...
04 March 2018 3,143 2 View
I know that chromosomal instability is responsible for triggering cellular death in M2, but what pathway or signal is activated to connect the DNA damage and subsequently trigger apoptosis? Does...
03 March 2018 4,656 2 View
Hi all, When I am conducting PCR in order to amplify a certain DNA segment for subsequent Sanger sequencing, would using the conventional DreamTaq Green Mix from Thermo Fisher work fine? Or...
13 February 2018 7,520 3 View
Hello all, I am performing telomerase inhibition experiments that will result in replicative senescence in cancer cell culture. Can anyone recommend assays to use to measure replicative...
05 February 2018 8,938 2 View
I treated wild type A549 cells with 10 nM RITA, a p53 activator drug, and the cell viability increased 18.7% in a statistically significant manner (p
10 January 2018 8,054 1 View
I wanted to know if anybody has ever done MTT Assay outside of a 96 well plate before, possibly inside a larger container by increasing the volume of reagents used?
19 September 2017 6,394 5 View
Trying to save cost here...can I grow tissue culture in a non-CO2 incubator as long as the media is supplemented with HEPES as a buffer instead? Thanks.
31 July 2017 2,400 3 View
I'm working with a DNA purification kit in order to purify and collect plasmid DNA from E. coli bacteria. The manufacturer's instructions say to resuspend a bacterial pellet formed after...
23 July 2017 5,769 4 View
Hello everyone! If I was administering CRISPR Cas-9 in vivo, and I wanted it to act only upon tumor cells within a certain region and not normal cells, how could I go about doing that? Would it be...
23 June 2017 5,161 2 View
Will the Cas-9 enzyme still be able to recognize and induce double strand breaks on condensed or highly condensed regions of DNA within eukaryotic cells?
22 June 2017 494 2 View
Normally guide RNAs have been 20 nucleotides long, but I am wondering if I can select to make them longer in order to reduce off-target activity. For example, say there is a gene of interest which...
22 June 2017 1,139 5 View
I am aware that there are two different methods for double strand break repair (non-homologous end joining or homology directed repair), but what I wanted to know is if the repair mechanism...
19 June 2017 5,694 3 View
I want to subculture a 75 cm^2 flask of MCF7 cells, grown in standard conditions (37 degrees C, 5% CO2), into a subdivision ratio of 1:4. How long does it take to culture these cells before they...
13 December 2016 6,759 4 View
I have heard worked with assays such as the MTT assay to measure cell viability, and heard of caspase assay kits to measure apoptotic activity. Yet what other methods and assays exist to measure...
13 December 2016 137 6 View
