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Concentration question

protein RNA AMPPNP

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#1 itmaybe

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Posted 08 June 2015 - 12:23 AM

Hello everyone,

 

I need help about something really simple but it makes me wonder (for some reason smile.png

Ok, I am doing binding assay of protein and RNA, I use ATP in the mixture (25mM). Usually I use 2.5 ul of that ATP in mixture which is together with buffer total 40 ul (200ng/ul of RNA and protein each beginning concentration).

Now, my question is: if I use higher concentration of RNA and protein (for example total 10 ug) and total volume is 100 ul, how much of 25 mM ATP do I need in this mixture?

 

Thanks for the help!!


Edited by itmaybe, 08 June 2015 - 12:26 AM.


#2 mdfenko

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Posted 08 June 2015 - 03:48 AM

if you are using the same final concentration of atp then just multiply by the volume increase (100/40=2.5x)


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#3 itmaybe

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Posted 08 June 2015 - 04:00 AM

Thank you very much for your reply.

I was just wondering does higher concentration of RNA and protein require more ATP...

Ok, thanks!! biggrin.png


Edited by itmaybe, 08 June 2015 - 04:05 AM.






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