I am trying to analyse RNA synthesis and obtain pulse-chase upon silencing of my GOI. I would like to obtain RNA content from the cells under different time courses which will be measured via FACS and Immunofluorescence simultaneously.
Any suggestions will be highly appreciated. Thanks in advance.
Hi Rabiul,
So far as I know, BrdU is used to incorporate into newly synthesized DNA when cells enter and progress through the S phase of the cell cycle. There are so many commercial BrdU Flow Kits available. I would recommend you the kit from BD company.
Actually, people add BrdU chemicals into the cells directly. It's quite a simple way to do it. However, you may need to optimize the time point and pulse-labeling time intervals for your cells.
Hope this information is helpful for you. Good luck!
Thank you so much. Indeed, knowledgeable information. I am quite familiar with Brd-U labeling of different cells including cardiomyocytes of my interest. I am seeking BrU labeling to analysee RNA stability via pulse-chase.. which can be alternative of usage of intercalator like actinomycin D.... https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3583853/pdf/rna-9-1233.pdf
Thanks again.
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