- Seed HeLa cells microporated with 5μM MBs so that they will
be 10–30% confluent at the time of experiment. - Dilute the target RNA stock to a concentration of 20μMin
Microinjection buffer. - Centrifuge the target RNA sample for 20 min at 21,000g.
Use the supernatant for injection as the pelleted debris can clog
the microinjection capillary.
Fig. 2Nonspecific opening of non-PS-modified and PS-modified anti-luciferase MBs in living cells. (a)
Representative images of MBs in HeLa cells, acquired at 10 h following microporation. The MBs are not
complementary to any known endogenous RNAs. Theinsetshows an expanded segment of the image. The
arrowpoints to a bright punctum in the nucleus. (b) Quantification of the extent of MB nonspecific opening over
time. Each data point represents the meanstandard error from at least 40 cells (scale bar, 10μm)
(Reproduced from [19] with permission from Elsevier)
Optimizing Molecular Beacons 251