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Gene copy number by slot-blot hybridization

David Barrass david at barrass1.no.spam.ta
Thu Jan 27 04:49:29 EST 2000


On Thu, 27 Jan 2000 00:20:19 +0800, lumdicks <lumdicks at netvigator.com>
wrote:

>       I'm planning to use slot blot hybridization to calculate the gene
> copy number in the genome. However, my slot blot give a very weak signal
> even 50ug of genomic DNA was applied. Is there any possible casue for
> this..?? I am using DIG labelling and chemiluminecence detection. The
> system works fine for southern blot (using the same probe and genomic
> DNA), which can detect a single copy gene using 5 - 10ug of genomic DNA.
> So, I don't know what's wrong with the slot blot. Can anyone give any
> suggestion??

I battled with the same problem trying to determine copy number in
transgenic animals.  

Let me guess are you looking at mammalian genome?   If so I suspect
that this approach will not work :-=(.  There is too much negative DNA
in the way with large genomes.; the probe simply cannot find the
target.  I eventually concluded that Southerns were the only way until
I could persuade someone to buy me a Quantitative PCR machine.

If you do find a solution please post it, it would be very useful

David

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