Hi everyone. I am doing biochemical detection of DNA damage, using phenol/chloroform/isoamlyalcohol ( in another word, DNA fragmentation assay). But, unfortunatelly, no result. The drug I am testing I already checked by western blot, and cytochrome C assay, and it cause apoptosis. I have quite big pellet after extraction, but on visualization-nothing! I can see genomic DNA, and small fragments, i think RNA...
If someone has experience in this matter, please, advice me... Thank you!
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#2
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Posted 03 May 2010 - 07:59 AM
pola, on May 3 2010, 08:56 AM, said:
Hi everyone. I am doing biochemical detection of DNA damage, using phenol/chloroform/isoamlyalcohol ( in another word, DNA fragmentation assay). But, unfortunatelly, no result. The drug I am testing I already checked by western blot, and cytochrome C assay, and it cause apoptosis. I have quite big pellet after extraction, but on visualization-nothing! I can see genomic DNA, and small fragments, i think RNA...
If someone has experience in this matter, please, advice me... Thank you!
If someone has experience in this matter, please, advice me... Thank you!
to make it clear: you expect to detect DNA fragmentation caused by apoptosis; and you are sure that apoptosis happens; do you have checked the time course of apoptosis?
#3
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Posted 07 May 2010 - 01:42 AM
Yes, I am expecting that drug I am testing cause apoptosis. By western blot we could detect the activation of caspase 9 and 3, and cytochrome C at 24 hours. The fragmentation assay was performed at 12 and 24 hours after drug adding... I wonder if it is enough time
Thank for your suggestions.
Thank for your suggestions.
