Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Biology

elimination of Protein interference from DNA - for RT-PCR (Feb/06/2011 )

hi
i am trying to detected DNA in a protein solution of high concentration (approx. 10mg/ml) . i am facing accuarcy issue, in terms of over-estimation. the main suspect is Protein.
could you plz suggest me techniques for elimation of this.

thanks a lot

-rick112-

rick112 on Mon Feb 7 06:12:45 2011 said:


hi
i am trying to detected DNA in a protein solution of high concentration (approx. 10mg/ml) . i am facing accuarcy issue, in terms of over-estimation. the main suspect is Protein.
could you plz suggest me techniques for elimation of this.

thanks a lot

Hola, you donīt say what type of DNA you want isolate, I suposse genomic, so any of the kits for that with colum washes and elution could help you. Beforehand, extractions with phenol /isoamilic alcohol were used, but in both methods you loose the protein. If you are interested look your DNA purification suppliers or any Mol. Gen. manual. but wait more indications of the forum experts. Buena suerte

-protolder-

You should be able to remove the protein by chloroform extraction (or by running through a DNA extraction kit). You can't easily estimate DNA content in protein solutions by 260/280 measurement, though you may be able to using a fluorescent based dye.

-bob1-