Microinjections - hints?

Graham Atherton grggta at picr.cr.man.ac.uk
Tue May 16 05:53:25 EST 1995


I am attempting to microinject an adherent cell line with antibodies to
specific proteins that we want to 'knock out'. Initial experiments with
Bgal DNA constructs as a control and washing the DNA through Centricon
'sieves', resuspending in PBS gave poor results. One cell stained up
blue the following day out of 60-70 attempted injections. 
  For the second experiment I used distilled H2O as solvent and got better
results - three blue cells out of 25 injected plus another 5 cells with
small blue specks at the site of injection (perhaps the needle was
blocked?)
   We really need higher success rate than this, has anyone any
experiance of successful microinjecting, or could anyone put me in touch
with someone with good experience? - please mail me so I could go into
more detail.
NB I am using a semi-automatic Zeiss injection system.

Many thanks
Graham Atherton
grggta at picr.cr.man.ac.uk



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