No, I have 1ug/uL DNase I and want to dilute it 1:2500 in the cold reaction 1X buffer. I only have a 10X stock. (This is where I got the 1/10).
Basically, I'm trying to find out how much DNase I to use, along with how much 10X reaction buffer to use to get my DNase I to dilute 1:2500 in a 1X reaction buffer.
What I did was (1/10)* 2500 = 250 fold dilution
I think this would give me the dilution factor for 1x buffer. With this dilution factor, I'm a bit uncertain where to go from here. My DNase I weighs 1 ug/uL. I need to somehow dilute this DNase I down 1:2500 in a 1X reaction buffer.
So I have to make a 1X reaction buffer from my 10X reaction buffer, and I think I found the dilution factor for it so far. Then, I have to dilute down the DNase I with this buffer to give me a final 1:2500 dilution. Since 1ug = 1uL, then I did 1uL*250 = 250uL. Or should I divide instead (1uL/250)? Whichever way, my goal is to find how much DNase I to add into the 1X buffer.
Finally, how much of the reaction buffer do I need to add in? Do I subtract (amt of DNase I) from 2500?
I don't completely understand what your calculations are and I'd like to be exact. I hope this clears up what I'm asking.
Edited by claritylight, 24 March 2005 - 05:46 AM.