Fig. 5 ES Cell Transfer
Microinjection is carried out as follows:
An embryo is brought into the injection position using the holding
capillary. The tip of the injection capillary is aligned with the
pronucleus and inserted directly into the pronucleus. Approximately
1 pl DNA solution is injected during every injection process. The
injection can be considered successful when the pronucleus increases
in size. If the volume of the pronucleus does not increase, the injection
is considered unsuccessful. The injection would then occur into cytoplasm,
where the injection drop shows different contrast. If the tip comes
too close to the endogenous DNA during the injection process, the
DNA sticks to the capillary when it is pulled out. If this occurs,
the capillary should be discarded. The correct injection volume of
the Transjector is set by defining the injection time and the injection
volume for all embryos.
Femtotips and Microloaders
Femtotips, the standard capillaries for microinjection,
are ideal for the injection of DNA, RNA, proteins and dyes into living
cells. The shapes of the capillary tip ensures high cell viability
and minimizes blockages by the injection material. The capillary opening
is constant, allowing reproducible injection volumes in the femtoliter
range.
Femtotips are available in two shapes for injecting cultured cells
or suspended cells. Backfilling with an Eppendorf Microloader easily
'"/>
Source:
Page: All 1 2 3 4 5 6 7 8 9 Related biology technology :1.
Biolistic Gene Transfer to Generate Transgenic Schistosomes, Rev A2.
Transgenic Zebrafish by Electroporation3.
A Positive Selection Assay for Mutation Analysis in Big Blue
Animals