Dear Ranjan,

If I underdstand correctly you want to do digital PCR.

Why not use the Biorad digital PCR system, or use other systems like Wafergen or the Quantstudio from Thermofisher. You could even do Next Generation sequencing on an Ion Torrent plarfrom.

Martin.

Actually this project is regarding emulsion PCR. I know about digital PCR.

Ranjan,

Maybe read this publucation first and find out if this is what you are looking for:

https://www.ncbi.nlm.nih.gov/pubmed/25744265

But this is in different language probably in French which I don't understand. Thanks. Can you help?

You are correct, did not realize this.

Try this link:

http://www.acgs.uk.com/media/1014434/10.40_-_02_-diagnostic_ctdna_analysis_of_egfr_in_non-small_cell_lung_cancer_angharad_williams__cardiff_acgs_2016.pdf

Thanks but this ppt mentioned about the pros and cons of ddPCR. Not the methodology. My problem is probably related with physical chemistry. I want to make a stable water in oil emulsion for PCR.

I have been through the same problem

I tried to make the emulsion using tween80/span80/triton X-100

Emulsion was made, but when PCR cycle is started the emulsions were broken btw the oil/water phases as you said

At that time, I concluded that the composition of the paper did not work or not proper for PCR

Recently, I saw the paper that uses the same emulsion composition

Article Simple paired heavy- and light-chain antibody repertoire seq...

Have you solved this problem?

I really wanna know the solution

Thanks

Thanks Kim for the update. We are still working on the project. But as I mentioned, the compositions mentions in the papers did not work for us. So we tried different ratios. Try using 1:2 and 1:3 combinations of water and oil.

Thank you for reply,

You mean that the ratio of the mixture (water:oil) has to be adjusted to 1:2 or 1:3? (the reference papers used a ratio of 1:4 (water:oil))

Yes