Separate DNA and dgtp

[Luke]

Hi Guys

I have a solution with DNA-Strands and dgtp (or datp, dttp, dctp). How can I isolate the DNA-Strands? I will make an experiment in which you do this very often, so it should be easy and fast. For example to filtrate or something like that… Has anyone an idea?

Kind regards

Luke

[Mega [Andreas Stuermer]]

Miniprep colums do this, but usually are expenisve. Maybe you can re-use them after HCl treatment.
Fragments smaller than 10 bp or so are not kept

[Michael Tellier]

PCR purification kit (if your DNA is > 100 bp) or the Nucleotide Removal Kit (for more than 40 bp DNA) are clearly the quickest and most reliable way (Qiagen but other companies also sell this stuff, sometimes cheaper). The columns can be reused at least 15/20 times (might be even more but didn't try it). I put them in 1M HCl for a few days before washing them twice with water, then air dry them.

Another possibility is to run your DNA on agarose gel but it is more time consuming and still need to melt the agarose band containing your DNA. It is more clean if you want to purify PCR products since you can cut the band with the right size rather than retrieving all the DNA > 100 bp with the PCR purification kit.     

[SC]

Hi Luke,

 

A regular precipitation and spin will also accomplish this.  The DNA will be in the pellet, but the loose dNTPs will stay in solution. 

 

Also, depending on what you would like to do with the DNA (ex: cloning), it may not be necessary to remove the dNTPs.

 

[Mega [Andreas Stuermer]]

Good idea. Google ethanol precipitation of DNA

[Luke]

thanks guys, it worked :)