MCF7 cells seeded on cover glass were transfected with siRNAs for 48 h before fixing with fixation buffer (4% formaldehyde, 10% acetic acid, 1 × PBS) for 10 min. Cells were then permeabilized in 70% of ethanol overnight and rehydrated in 2 × SSC buffer (300 mM NaCl, 30 mM sodium citrate (pH7.0)) with 50% formamide. Hybridization was carried out in the presence of 30 ng of probe (Sangon Biotech, biotin-aaaGTAGTTGTGCTGCCCTGGCAATGATTTAGAC) at 37 °C overnight. Biotin-labeled probes were incubated with Streptavidin-Cy3™ (Sigma Aldrich, S6402) in 2 × SSC buffer with 8% formamide, 2 mM vanadyl-ribonucleoside complex, 0.2% RNase-free BSA at 37 °C for 1 h in dark. Nuclei were counterstained with DAPI (0.1 µg/mL) and then washed twice with 2 × SSC with 8% formamide at room temperature (RT) for 15 min. Three images of each cover glass were taken with Carl Zeiss laser confocal microscope, and representative images were shown.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.