Chem/Biochem guys

[TDiddy]

I'm trying to work through an issue for work - a high throughput genotyping lab. We are currently testing a new method of DNA extraction using a NaOH based extraction solution. Some of the tissue samples have a small coating of iron-based paint, and there is a question of whether this paint is affecting the downstream PCR reaction at all.

First sets of data seemed to indicate that the paint didn't have any detrimental effect, but now its being called in to question. Is there anything I can try adding to the extraction solution that will bind or chelate the iron but not interact with the DNA in solution? Bentonite maybe?

[oly884]

Well, my guess would be that bentonite would pull the DNA out of solution as well as the iron, that stuff is fairly unforgiving when it comes to pulling things out of solution.

I'd say take a look at orthophosphate. From some reading it appears that it's a good way of getting iron out of solution relatively quick.

If not that, take a look at some other iron precipitation reactions.

[oly884]

Here's a link talking about orthophosphate's effect on iron:

http://www.epa.gov/ordntrnt/ORD/NRMR...alconf2002.pdf

[bamachem]

sorry, Chem-E, but I just put beans in a can.

looks like Oly's on the right track to me.

[TDiddy]

Cool, I will check that out, thanks David. I've had success in other extractions using bentonite to pull out proteins and other junk, I just wasn't sure how it work with Fe.

[waskillywabbit]

Add some worchestershire.

[MTL_4runner]

Add some worchestershire.

I was waiting for an ME to chime in.

[TDiddy]

Add some worchestershire.

Oh, go AutoCAD something...

[Euphorickaty80]

Is this thread in english??

[oly884]

ChE's > all other engineers