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Ethanol precipitation of DNA

ethanol precipitation DNA nucleic acid

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21 replies to this topic

#16 pito

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Posted 05 June 2015 - 10:49 AM

btw: if you resuspend it, let it sit for a while, do not add the water/TE and measure the concentration right away, wait a few minutes, heat maybe, just let it "sit" a while then measure...

 

Normally its pretty straightforward.

 

I guess you just had some bad luck with this particular sample that gave a weird result.


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#17 mdfenko

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Posted 05 June 2015 - 10:49 AM

i did say it would be difficult to dissolve if over dried.

 

you can, maybe should, use te but it depends on what you plan to do with it next. you can also use tris without the edta.

 

pito is correct, the pure  dna pellet is virtually invisible. that's one of the reasons to add glycogen.


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#18 Mad Researcher

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Posted 05 June 2015 - 10:54 AM

Thanks, i will let you know perhaps on Monday what happened.

 

So, after the ethanol precipitation the 260/280 and 260/230 ratio should inc? will it affect the yield also?


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#19 pito

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Posted 05 June 2015 - 11:00 AM

Thanks, i will let you know perhaps on Monday what happened.

 

So, after the ethanol precipitation the 260/280 and 260/230 ratio should inc? will it affect the yield also?

 

should what? inc?

 

THe ratios will be a bit different. I am not sure, but I thinks its the 260/280 ratio that is going to be a bit higher..

One of them is a bit higher !

THe other one: a little bit higher, rather 2,1-2,2 than 1,8 , If I remember correctly.

 

The yield? Not sure what you mean... You will lose some DNA of course...
 


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#20 mdfenko

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Posted 05 June 2015 - 11:00 AM

yes. you will end up with less than you started.


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#21 Mad Researcher

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Posted 05 June 2015 - 11:02 AM

ok. That is what was confusing me. I just wanted to confirm the "yield won't ever improve" but the DNA would be purer than before.


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#22 pito

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Posted 05 June 2015 - 11:07 AM

EUm yield , I think you are mixing up terms. Most often, when using yield in this case, they mean a % or just how good the "yield" is, how much you have from the initial amount.

Of course: the DNA will not "become more", I mean: you start with X DNA, it can never become more.

 

The "yield" can be pretty good, but it will Always be less of course.

 

THe DNA purer than before? Depends on what you mean with "before"... You mean after a restriction digest or? Yeah, of course...

Normally you use the ethanol precipitation  indeed as a way to clean up your DNA so yeah, it should improve the cleanliness.

 

ok. That is what was confusing me. I just wanted to confirm the "yield won't ever improve" but the DNA would be purer than before.

 


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