heparin in DNA - no working PCR (Aug/12/2008 )

Hi everybody there,

does anybody of you can help me with this: I isolated DNA from blood taken to heparin tubes, didn´t realised, that heparin is strong inhibitor of PCR (I have no product at all). Is it possible to clean DNA from heparin? If so, how?

Thanx, Thanx, Thanx for help!

yana

There are a lot of potential methods to remove heparin.

There are non specific glycosidase and protease enzymes that can degrade the highly glycosylated heparin.

Also, boiling the DNA in heavy reducing buffers ...

Maybe Ph/ChCl4/IAA (Phenol, Chloroform) extraction will also remove much of the protein.

Any or all of these methods could reuce or eliminate protein/saccharide levels.

Heparin is H2O soluble "sugar" and phenol/chl cannot remove it. You may try reprecipitate the DNA in ethanol.

hello

treat your DNA with 5 U heparinase I to remove heparin residues, taken from :-


Heparin Inhibits DNA Synthesis and Gene Expression in
Alveolar Type II Cells.
Cheng-ming Li, Donna Newman, Jody Khosla, and Philip L. Sannes

Yee, thanks, I have ordered heparinase I...hopefully it will works...I am curious about.

Have a nice day all
y.

hello

treat your DNA with 5 U heparinase I to remove heparin residues, taken from :-


Heparin Inhibits DNA Synthesis and Gene Expression in
Alveolar Type II Cells.
Cheng-ming Li, Donna Newman, Jody Khosla, and Philip L. Sannes