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        Transfection protocol for 6-well dish

        互联网

        1807

        Seed cells at appropriate density (5x104/well for 3T3, MPAC, BHK; 5x105/well for aTC1.6, bTC3) one to two days before transfection.

        Combine total DNA at 1ug /well with 600ul/well serum-free medium in an eppendorf tube.

        Add 1.5ul/well Transfast (Promega) to the DNA/medium mixture. Vortex for 10 seconds.

        Incubate at RT for 10 to 15 min.

        During the above incubation, aspirate medium from cells and add PBS; remove PBS just before adding DNA in step 6.

        Add 600ul DNA mixture from step 3 to the appropriate well.

        Incubate at 37 oC incubator for 1 hr.

        Add 4ml regular growth medium to each well.

        after 24 hours, change to fresh medium.

        After 48 hrs, harvest cells for functional assay.

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