Manual: AdEasy Adenoviral Vector System - UCLA Human Genetics
Manual: AdEasy Adenoviral Vector System - UCLA Human Genetics
Manual: AdEasy Adenoviral Vector System - UCLA Human Genetics
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Growth Medium<br />
DMEM (containing 4.5 g/L glucose and<br />
110 mg/L sodium pyruvate and 4 mM<br />
L-glutamine), supplemented with 10% (v/v)<br />
heat-inactivated fetal bovine serum<br />
Growth Medium (supplemented<br />
with 25 μM chloroquine)<br />
Note<br />
Chloroquine solution is toxic and<br />
should be opened in the laminar<br />
flow hood<br />
Prepare growth medium as above. Add<br />
chloroquine from stock chloroquine solution<br />
to a final concentration of 25 μM. Filter<br />
sterilize. Prepare just before use and keep at<br />
37°C until required.<br />
Stock Chloroquine Solution<br />
(25 mM final concentration)<br />
Note<br />
Chloroquine is toxic and should be<br />
opened in a fume hood only<br />
1.29 g of chloroquine diphosphate<br />
[C18H26CIN3 • 2H3PO4]<br />
Add 100 ml of 1 × PBS; dissolve the solid<br />
chloroquine. Filter-sterilize and store in<br />
aliquots at -20°C. Discard aliquots that are<br />
older than one month. Dilute 1:1000 into<br />
media for use during the transfection<br />
MBS-Containing Medium<br />
Note<br />
Chloroquine solution is toxic and<br />
should be opened in the laminar<br />
flow hood<br />
Add stock chloroquine solution to DMEM<br />
containing 7% (v/v) modified bovine serum<br />
(Solution III from the ViraPack Transfection<br />
Kit) to a final concentration of 25 μM. Filter<br />
sterilize. Prepare just before use and keep at<br />
37°C until required.<br />
38 <strong>AdEasy</strong> <strong>Adenoviral</strong> <strong>Vector</strong> <strong>System</strong>
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