injection problems

Morris Maduro Morris_maduro at biology.ualberta.ca
Thu Oct 27 11:01:48 EST 1994


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



More information about the Celegans mailing list