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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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200bp fish probe
Posted by: fatboym (IP Hidden, Junior member, 20)
Date: March 12, 2005 11:07AM
I have no good experience in FISH and I would like to know if a 200bp probe will be visible on a parafin empbeded slide.
Re: 200bp fish probe
Posted by: sheantine (IP Hidden, Unregistered user, )
Date: March 13, 2005 06:39PM
200-bp is the opimal size for FISH probe in terms of cell penetration, specific activity and detection sensitivity. You are on the right track!!
Re: 200bp fish probe
Posted by: fatboym (IP Hidden, Junior member, 20)
Date: March 14, 2005 01:51PM
well, i have read about this, but then why the commercially used probes for gene amplification are at least 100kb long?
Re: 200bp fish probe
Posted by: fatboym (IP Hidden, Junior member, 20)
Date: March 14, 2005 01:51PM
oh. thank you very much for your interest and reply
Re: 200bp fish probe
Posted by: sheatine (IP Hidden, Unregistered user, )
Date: March 14, 2005 06:34PM
"at least 100-kb" ...."gene amplification", are you living on earth??? Are we speaking the same language??
Re: 200bp fish probe
Posted by: fatboym (IP Hidden, Junior member, 20)
Date: March 15, 2005 03:07PM
The LSI C-MYC (8q24.12-q24.13) Probe is an approximately 120 kb SpectrumOrange labeled probe.
Cyclin D1 (also known as PRAD1 and CCND1), probe may be used to deter-mine the copy number of the Cyclin D1 locus, or as an enumerator probe for chromosome 11 in interphase and metaphase cells. The LSI Cyclin D1 is approximately 300 kb in size. I copy here sentences from the Vysis site. Iknow from literature that 200bp is the optimal size for a probe in order to enter efficiently in a parafin-embeded tissue. However, I have no experience of my own what kind of signal such a probe is going to show me, and I haven't found anywhere pictures or articles from FISH with such a small probe. So I am afraid that it is too short to work. I may be fool, but I would be very interested in picures or examples of FISH with probes around 200bp.
Re: 200bp fish probe
Posted by: sheatine (IP Hidden, Unregistered user, )
Date: March 15, 2005 04:44PM
You're talking about 2 quite different things here - chromosome FISH and cellular/tissue FISH.
In your previous email you mentioned FISH application in parafin-embeded tissue, while you used chromosome FISH as examples here to show 100kb probe??????? This is not a good way to prove your point, totally non-scientific????? You can easily find tissue FISH picture using 200 bp probe on internet, be patient!!! I would be extremely astonished if you can use 100 Kb probe for tissue FISH, but love to see it.
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