Q: Can you random prime a 250bp PCR product?

[Heinz J. Schaefers]

Hi Makato,

i have tried to random label a pcr fragment of 305 bp for a northern probe,
but all i got was unspecific binding and high background on the northern blot.

After cloning the fragment in a TA cloning vector, and cutting it out again,
and random labelling, the probe i got was fine, no background, and no
unspecific bands.

Heinz

Makoto Kamei wrote:

> Hi all,
>
> I am about to screen a mouse genomic DNA library constructed in EMBL3
> lambda vector. I have ~250bp PCR product made from mouse genomic DNA
> template. Is it possible to use this product as a probe for the screening?
> Is it possible to random prime this product to make a probe? Any help
> appreciated. Thanks a lot in advance.
>
> Cheers,
>
> Makoto Kamei
> Makoto.Kamei at anu.edu.au