probe size in FISH
liuz at omrf.ouhsc.edu
Mon Apr 3 08:57:22 EST 2000
I currently want to use the method of fluorescence in situ hybridization to
locate which allele of the pair of chromosomes of ES cell was targeted by my
targeting vector because I had a heterozygote of ES cell, in which one
gene had been targeted, and I have to know if my new targeting event
occurred in the same chromosome or not. In my pre-experiment, I tried two
probes on metaphase chromosome spreads. Both should be positive because
they have the same origin with chromosome. I tried once, but failed without
signals except high background. The probe size I used is that one is about
and the other is about 0.6kb. I cloned both of them in cloning vectors and
labeled them using the whole plasmid (insert + Vector). Before I try
questions are below.
1. How long is the minimum size of a probe applied in FISH on metaphase
chromosome? Does 0.6kb work well?
2. How can I reduce the background? Don't forget I use mouse chromosome.
3. Is there a good book for all aspects of FISH?
My e-mail is liu at omrf.ouhsc.edu.
Thanks a lot.
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