Does anyone have a protocol or know of a publication that describes the PCR along with the primers that can be used to differentiate mouse and human cells? I'm working with both and would like to confirm my cultures to ensure no cross contamination.
Thanks
Differentiating mouse from human cells with PCR
Started by Epigeneticist, Jan 16 2013 10:34 AM
PCR Cell Culture Genotyping Mouse Human
5 replies to this topic
#1
Posted 16 January 2013 - 10:34 AM
#2
Posted 21 January 2013 - 05:07 PM
Depending on the cell lines you are using just find a mouse-specific or a human-specific gene. Go to pubmed, find gene and design primers.
You can't do a visual check to see if your cells are contaminated? Not the best method, but I don't understand why you are doing this.
You can't do a visual check to see if your cells are contaminated? Not the best method, but I don't understand why you are doing this.
#3
Posted 22 January 2013 - 05:59 PM
Jerry Shelly 1
I am aware of this option to find a gene and get some primers. However, I was wondering if anyone uses this method and if there are some popular regions to amplify. I'm not an expert on cell culture. Also, I do not have an issue with contamination at the moment but thought it would be a good way to do a confirmation of the cell culture. Thanks for the suggestions and any future ones you can provide.
I am aware of this option to find a gene and get some primers. However, I was wondering if anyone uses this method and if there are some popular regions to amplify. I'm not an expert on cell culture. Also, I do not have an issue with contamination at the moment but thought it would be a good way to do a confirmation of the cell culture. Thanks for the suggestions and any future ones you can provide.
#4
Posted 23 January 2013 - 10:33 AM
Which cell lines are you working with in both human and mouse.
#5
Posted 05 February 2013 - 07:00 PM
jerryshelly1, on 23 January 2013 - 10:33 AM, said:
Which cell lines are you working with in both human and mouse.
jerryshell1 - I think I have it figured out. However, to follow-up your question. I am not working with a human cell line but others in the lab are working with many different human cell lines. I am working with mouse primary cells.
Thanks for the help!
#6
Posted 22 February 2013 - 06:29 AM
We have had also problems with mouse cells contaminating human cells. In addition, we obtained once a mouse cell line that turned out to be a dog cell line. Since then we regularily check our cell lines for interspecies contaminations. We found a service lab called Multiplexion to do it at a very reasonable price. At the same time they also check for SMRV and Mycoplasma.
Also tagged with one or more of these keywords: PCR, Cell Culture, Genotyping, Mouse, Human
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