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what primers should I use for DNA sequencing? please help


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#1 springstreet

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Posted 19 October 2009 - 05:42 AM

I only know the vector is pED-3d, but dunno the insertion site. Now I need to submit samples for DNA sequencing, please let me know what primers should I use? (T7? or something else) If I have to make primers myself (without knowing the insertion sites), how should I do it?

Thank you all so so much!!

#2 microgirl

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Posted 19 October 2009 - 07:13 AM

I can't find a pED-3D vector but there is a pET-3D vector - is that it? Why don't you know your insertion site? Looking at the vector map, you could try T7 but if that's not near where your insertion is it won't do you any good. Otherwise I guess you could get the vector sequence and design some primers at various points around the vector near where insertion sites could be and just keep trying.

#3 springstreet

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Posted 19 October 2009 - 02:09 PM

View Postmicrogirl, on Oct 19 2009, 11:13 AM, said:

I can't find a pED-3D vector but there is a pET-3D vector - is that it? Why don't you know your insertion site? Looking at the vector map, you could try T7 but if that's not near where your insertion is it won't do you any good. Otherwise I guess you could get the vector sequence and design some primers at various points around the vector near where insertion sites could be and just keep trying.

thanks, I will try to find more info on the insertion site.....




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