Hello again,
I have constructed a series of lacZ fusion and GFP fusions containing the
first half of unc-119. I have injected these plasmids, at various
concentrations, along with pRF4 into N2 animals. Of over 110 animals
injected I have seen no F1 rolling animals. When I injected pRF4 along
with a 3-kB genomic clone from outside the region, however, I obtained 10
F1 rollers (from at least 5 different parents) after injecting only 20
animals, suggesting my injection technique is not a fault. There are no
dead eggs on the plates where I tried lacZ and GFP fusion injections,
suggesting the constructs themselves aren't lethal. The only difference
between the preps of DNA is that I RNAse treated the plasmids in the lacZ
and GFP fusion cases, but not the genomic clone case.
Does leftover RNase in a plasmid preparation adversely affect recovery of
transgenic animals after microinjection? Have others had problems with
such lacZ constructions? Or am I just having extremely bad luck?
Frustrated,
Morris M.
--
Morris Maduro | 'Worms Rule'
Dept of Biological Sciences | 'OK'
University of Alberta |
Edmonton, AB Canada | Morris_maduro at biology.ualberta.ca
