[Zbrafish] transcription of probes

sunshyne61987 from yahoo.com via zbrafish%40net.bio.net (by sunshyne61987 from yahoo.com)
Tue Jul 3 10:40:59 EST 2007

Hi, I am trying to set up some in situ experiments using ntl as a
control along with 2 sequences of interest. We have run our cut
plasmids on a gel and they all look like distinct single bands. We
then made our probes with the Roche DIG kit and the ntl seem to give
us nice distinct bands, though the antisence is weaker. However, our
sequences give oly weak single bands and only when the plasmid is cut
with Xho and T3 is used (sense of one and antisence of the other). the
plasmids cut with Xba and in which T7 is used give us almost no
detectable bands. the surprising thing is that both the ntl and our
sequences are in the same Bluskript backbone vector. The problem seems
to be in the transcription of  our sequences....please help!

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