General Category > 2-D Electrophoresis

How long will it take to do 100 proteins identification and analyse pathways?

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yudohaoxiu:
Hello experienced people:

I am writing a proposal and need to estimate money/time etc., so I would be very appreciated to get some help from you people.

I did DIGE analysis in my PhD and found about 100 proteins that have up or down regulated significantly, and now we are interested to get those proteins identified by running a preparative gel, and hopefully find some pathways that is interesting. As I have never done this before, I would like to know how long does it normally take? I would say if everything goes well, it would take 2 months (2 weeks culturing the cells, 2 weeks processing the extracted protein and the rest DIGE, Obitrap MS) to get those protein identified. Then I should look up those identified proteins in publications and see if there is any correlations between them. This seems to me another month work. If the result is good, we will be happy to make it into a publication. So I assume these can be done in 4 months?
I am planing to submit the cut out proteins (silver stained after CyDye labelled) to an MS service. I have run gels before so I am not very worried about that. The things that are new to me are spot picking (using the Ettan) and digestion. I wonder if these parts are tricky to have mistakes occurred or straight forward? I know these are highly related to the skills of different people but I really want to get some information. 

Thank you for your patients. I just want to get a good picture before I talk to my boss.

Jenny

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