Carrier DNA inhibits transfection

Dr.Sailesh Surapureddi SaiSu at MCB.LiU.SE
Fri Jan 5 14:52:22 EST 1996


>To: methods at net.bio.net
>From: david_levy at dfci.harvard.edu (David N. Levy)
>Subject: Carrier DNA inhibits transfection
>Date: 5 Jan 1996 17:18:24 GMT
>
>I have recently tried using sheared salmon sperm DNA (Sigma, 
>sodium
>salt) as a carrier in calcium phosphate-BES trasfection if Vero 
>cells
>and have found that it efficiently inhibits transfection of 
>various
>luciferase reporter plasmids.  My shearing looks good on an 
>agarose
>gel.  Is there a contaminant present that inhibits transfection?  
>I
>have not tried boiling the DNA yet.  Any experience with this 
>problem? 
>Thanks
>
>
>David N. Levy
>Division of Molecular Genetics
>Dana-Farber Cancer Insitute
>Boston, MA 02115
>david_levy at dfci.harvard.edu


The phenolic (for that matter even Ethanol) residue left over in 
the preparation of SS-DNA, kills the transformations and 
transfections. 
High boiling usually helps in evaporation of Cholorom and 
Ethanoic contaminations. so it might help you actually.
Cheers
Sailesh.



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