Hi,
Today I want to ask about stability of cDNA and protein.
I often conduct RT-PCR and Western blotting, starting from huvec culture.
During RT-PCR / Western blotting,
I wonder whether such happenigs below can occur
by unnoticed factors or by my mistakes.
1) cDNAs originated from huvec can change into cDNAs originated from
other types of cells?
2) Proteins originated from huvec can change into proteins originated from other types of cells?
These may be consisent with my previous question about my concern of
change of cell type.
I think my question above may sound ridiculous and obsolete but
I am yet an beginner in research so I want you to understand my
current state.
Thank you!
Once you have purified protein or nucleic acid it is overwhelmingly unlikely (this is science, so I can't quite say "impossible") that your sample will change its composition to the point of seeming like it's from a "different type of cell." Though, that's hard to define as well.
Some things that COULD happen:
- Protein/nucleic acid degradation. In this situation, it may artifactually seem like a different isoform is purified, rather than the true form.
- Protein/nucleic acid aggregation. Same interpretation result as above.
- Protein/nucleic acid conformational change. Same interpretation result as above.
This is different than changing to be like it is from a "different type of cell" because the 1st order sequence of the sample is basically the same.
Dear Jordan R. Yaron,
I appreciate your informative and structured answer!
I learned a lot thanks to your help.
Hope for great achivements in your research!
Dear Lee
if there is a problem with cDNA from other cell types then the simplest explanation to posit would be contamination of your cell population by something else culminating in cDNA from 2 or more sources. I cannot really add to that or indeed the above
- Badges
- Science method