a breast cancer cell line, MCF7, was used for an in vitro study to investigate the therapeutic potential of a drug. DNA fragmentation kit (Apoptotic DNA ladder kit from Roche) protocol was followed to assess the change at the molecular level upon drug treatment.
after running the gel for DNA ladder visualization, both - control ( untreated cell) and the treated cells show similar banding pattern, check the attached file.
Ideally, the non apoptotic (- control) should display one intact band, where the apoptotic cell show a fragmented smear as indicated in the attached gel pic (+ control).
Your assistance and feedback are highly appreciated to interpretation and troubleshooting.
Dear widyan,before the running of treated and control cell DNA, you have to run the control only.if there were one intect band in all control then thats were ok.but if there were any fragments that was happened due to contamination of RNA.so you have to use specific amount of RNAase enzyme to remove RNA.
I agree with Artur about apoptosis that has been established as a conserved mechanism over decades beyond any doubt etc. For the gel, I also agree there is a lot of RNA -the main ribosomal species are obvious. Most of the DNA is stuck on top-in the loading well and therefore have high MW to enter s the gel. No signs of degradation are seen. Wharever was used to induce apoptosis di not work
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