Ori Elad

12 Questions 15 Answers 0 Followers

Questions related from Ori Elad

Inconsistencies in PCR results after bisulfite conversion?

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 4,805 1 View

Should there be a correlation between band size on gel and SYBR green signal in the real time PCR run?

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 9,033 4 View

Is there logic behind using Annexin V FACS with another antibody?

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 7,848 3 View

What are the alternatives to 5-azacytidine for bursal B cell cultures?

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 4,731 3 View

High miniprep yield followed by extremely low Ct values?

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 1,032 7 View

How do I use two way ANOVA on real time PCR results?

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 5,304 7 View

How do I calculate standard errors for normalized log2 converted gene expression data?

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 8,782 7 View

Has anyone had any experience with storing cell culture slides for a TUNEL assay?

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 3,575 2 View

Can anyone recommend a process for preparing and storing slides for a TUNEL assay?

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 4,811 2 View

Does anyone have a specific recommendation for a DNA pol or mix fit for PCR ?

I'm using bisulfite treated DNA as template for sequencing purposes.

09 September 2014 6,797 7 View

Any problems using the same primer set twice?

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 5,412 8 View

What are acceptable concentrations of Folic acid and 5-azac in DT-40 cell culture?

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 1,031 2 View