[Zbrafish] probe synthesis problems?

sarah at RIC via zbrafish%40net.bio.net (by sspinette from ric.edu)
Thu Jun 28 12:42:37 EST 2007


We are starting up some in situ expereiments. We are using a Ntl
plasmid as a control along with our sequence of interest (which is
about 750nts). We ran some of our antisence and sence probes on a gel.
The ntl probes both look like beautiful sharp bands of the right size
with no degredation detected. However, in the lanes in which we ran
our probes of interest we see a faint band of the right size but we
also see a bunch of larger bands as well ranging from ~1000nt-2500nt.
we checked that our plasmids ere fully linearized with the right
enzyme. We are wondering if it could be secondary structure that is
resistant to 65 deg incubation or some other problem...please help!



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