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Questions related from Ori Elad
Hello all. Around two years ago I extracted DNA using a qiagen kit from a DT40 cell line culture. Yield was around 100 ng/ul with a 260/280 ratio around 2. I then used the epitect kit to treat...
04 April 2016 4,820 1 View
Hello all. I have been working with a sequence that is a 571 bp clone, and I have a primer pair that gives a 150 bp amplicon. When I ran the system in real time for a calibration curve I got some...
01 January 2016 9,050 4 View
Good evening. I am wondering if it is possible to do an Annexin V FACS staining on B cells extracted from the bursa together with staining with an antibody against surface IgM. I read that B...
12 December 2015 7,860 3 View
Hello all. B cells isolated from chick embryos' bursa do not survive in culture for very long. (down to 30% viability after 9 hours of incubation). I really wanted to test the effect of...
11 November 2015 4,744 3 View
I have been using Clone jet PCR cloning kit with a pJET1.2 blunt as a vector. The insert is from a clean PCR product that went through a blunting reaction (supplied with the cloning kit). The E....
10 October 2015 1,047 7 View
Hello all. I want to test whether two different factors - concentration (3 treatments) and incubation time (2 treatments) have an effect on gene expression. I ran the gene, and I have the Ct...
03 March 2015 5,321 7 View
I ran an experiment with a ref gene (3 biological repeats x 3 technical repeats) and normalized gene expression of my target genes to the ref gene. I decided to present the data as log2 fold...
01 January 2015 8,792 7 View
Hello all. I have read a protocol stating that post paraformaldehyde fixation (4% in PBS) cell culture slides can be preserved in 70% ethanol at -20 deg. cel. until use (not good for long term,...
12 December 2014 3,591 2 View
I want to prepare slides of DT-40 cells for a TUNEL assay. My question is, is there a protocol for preparing them and fixing them so they can be stored at -80 degrees for about a week?
09 September 2014 4,825 2 View
I'm using bisulfite treated DNA as template for sequencing purposes.
09 September 2014 6,810 7 View
Theoretically, is there a problem with running a PCR and then running the same system again, with the same primers, with the product of the first reaction to further amplify the product? The...
08 August 2014 5,424 8 View
I'm working on Methylation/Demethylation of certain promoter regions in DT-40 cell genome. Any recommendations as to folic acid/5-azac concentration and exposure time for that type of cell...
05 May 2014 1,042 2 View
