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DNAseI question

DNAseI

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#1 newtogenetics

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Posted 15 February 2012 - 02:04 PM

Hi all!
I am a very new lab tech with little training, so this question will most likely seem a little silly to most:

I am using a DNA extraction protocol which requires the addition 20 ul of 30ug/ml DNAseI (Roche) to 750ul of template. (then 30min incubation at 37C)

I ordered DNAseI and it came with a concentration of 1u/ul (1000units), and also came with reaction buffer and EDTA.

I've been reading about DNAseI and its use and such, but am still confused as to how get the DNAseI to be a concentration of 30ug/ml from 1u/ul. Am I supposed to be mixing the DNAseI with the reaction buffer to make it a concentration of 30ug/ml?

Like I said, kind of a silly question, but all of this stuff is so new to me!

Thanks for any help.

Edited by newtogenetics, 15 February 2012 - 02:35 PM.


#2 dimitris31b

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Posted 16 February 2012 - 05:15 AM

Why use DNAse on DNA!!!! It will destroy your DNA. Do you mean RNAse instead?

#3 HOYAJM

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Posted 16 February 2012 - 08:33 AM

Yes, for DNA extraction you want RNAse. But, here is a link that should help you convert U/ul to ug/ml

http://www.protocol-...osts/18064.html

#4 newtogenetics

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Posted 18 February 2012 - 02:56 PM

Thanks for your help! It actually is DNAse that I want (very weird, I know). I am extracting an endosymbiotic bacteria from its host, and the DNAse step is required for digesting host cells, before extracting the endosymbiont DNA.

#5 bob1

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Posted 18 February 2012 - 04:09 PM

Why use DNAse on DNA!!!! It will destroy your DNA. Do you mean RNAse instead?

Not necessarily true, it is common practice to have a DNase step before cell lysis to remove contaminating DNA.





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