We are looking to purify an excess of mammalian DNA away from
bacterial DNA as applied to a tissue extraction. The reason being that
the DNA needs to be concentrated up before a detection PCR specific to
the microbe can work. However, at these higher concentrations, PCR
product visualisation on an agarose gel is obscured, because of
excessive mammalian DNA. In short, we are looking to bind a Brucella
specific probe onto capture beads, enabling concentration of Brucella DNA and elution of mammalian DNA. Is there anyone that can accommodate this intended
protocol, or has familiarity with it in a different context?

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Hi, you might want to try out a method used in this paper: J Virol Methods. 2003 May;109(2):209-16. Ultra-sensitive and specific detection of porcine endogenous retrovirus (PERV) using a sequence-capture real-time PCR approach. By Shah CA, Boni J, Bisset LR, Seebach JD, Schupbach J.

They used magnetic bead sequence capture approach to concentrate an endogenous retrovirus DNA. Oligonucleotides to a conserved gag sequence was attached to magnetic beads, these beads where used to concentrate PERV DNA from their sample material for PCR experiments.

Original Message
From: madscientist
Date: Friday, July 11, 2003 4:15:25 PM
I know that there are a couple of companies that make streptavidin coated magnetic beads (Dynal and Novagen both do) - theoretically you could biotin label a Brucella specific probe, bind it to the beads, incubate the probe/beads with your DNA mix and wash away the non-specific genomic DNA...

Original Message
From: Terry Murphy
Date: Friday, July 11, 2003 1:52:01 AM
We are looking to purify an excess of mammalian DNA away from
bacterial DNA as applied to a tissue extraction. The reason being that
the DNA needs to be concentrated up before a detection PCR specific to
the microbe can work. However, at these higher concentrations, PCR
product visualisation on an agarose gel is obscured, because of
excessive mammalian DNA. In short, we are looking to bind a Brucella
specific probe onto capture beads, enabling concentration of Brucella DNA and elution of mammalian DNA. Is there anyone that can accommodate this intended
protocol, or has familiarity with it in a different context?

Post ID: 1426 Link Reply Quote Report: Spam or Abuse