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who knows a protocol of a "making of a specific mouse probe"?
Prerequistes:
DNA probe (no chromosome probes), mouse specific (no cross reaction with human), existence in the liver.
Thanks
Tiffy
Mon Jan 10, 2005 5:12 pm
genedetect
Rank: Member
Joined: Oct 14, 2004
Posts: 85
Post subject:
Hi Tiffy
We have made a few of these types of probes for our customers with good results so far. Our approach would be to select (in this case) Mouse-specific satellite repeat sequences to target probe design to.
The result is a probe that only detects Mouse tissue and not Human tissue for example. I assume you might be performing some sort of tissue or cell implantation into Human and want to identify Mouse tissues from the surrounding Human tissue? If so, you might consider using this approach rather than try detect a specific Mouse gene.
Satellite repeat sequences can be found that are species-specific and since they are highly repeated in the genome this means lots of probe binding sites in the tissue and thus a nice strong signal.
thank you for nice offer. But the point is that I have already a very good functioning mouse probe, which was a gift from an another scientist. Because this was only a gift I asked for a protocol to create a probe on my own to be more independent.
You were right regarding the transplantation.
Best wishes
Tiffy
Tue Jan 11, 2005 10:47 am
genedetect
Rank: Member
Joined: Oct 14, 2004
Posts: 85
Post subject:
Hi
My advice then is to target Mouse satellite repeat sequences that you can find in Genbank. Then you screen different probes against the target repeat sequence (using any one of the common primer design programs) to get the one that has the best hybridization parameters and is devoid of secondary structures. Lastly, you check that the selected probe sequence does not detect any Human sequences by BLAST or Fasta.
By targeting satellite repeats you ensure you will get a very strong signal with the probe.
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