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4. The transfection/electroporation effi cacy can be monitored
for constructs with reporter genes by fl uorescence (GFP) and
X-gal staining (
-gal).
5. Generally, it is suffi cient to fi lter-sterilize the SFV stocks and no
further purifi cation or concentration is necessary. However,
the medium from BHK-21 cells has demonstrated some toxicity
to primary cells in culture. For instance, for studies in primary
neurons, it is recommended to subject the SFV stocks to
ultracentrifugation or affi nity column purifi cation.
6. Use only fresh batches (or aliquots) of
β
-chymotrypsin for the
SFV particle activation. Also notice that there is difference in
potency of different batches of
α
-chymotrypsin. It is advis-
able to include a reporter gene SFV stocks with known titers as
a control.
α
References
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