I have to run qPCR on RNA from mucosa from a pig's stomach. I have now tried 2 times without success. My RNA concentrations are too low to make cDNA. I get my stomach from a butcher 40 minutes away. The stomach is not cut up or cleaned and it is transported in a 37 degree hot krebs solution together with an air pump. Can the RNA degrade because of the stomach contents and lack of oxygen? Do you think it will help to cut the stomach open at the butcher and then put it in the krebs solution, so that it will touch the mucosa, or do you think it has no effect on the RNA?
I put a response in on the "discussion" thread of this same topic. I'm curious to see what you may think of my suggested approach.
Melissa Bemrose I'm new to this website, so I don't know where the "discussion" thread is, can you help?
Amalie Storm ,
This is the discussion to which I was referring:
https://www.researchgate.net/post/I_have_to_run_qPCR_on_RNA_from_mucosa_from_a_pigs_stomach_I_have_now_tried_2_times_without_success_My_RNA_concentrations_are_too_low_to_make_cDNA
I'm not sure why some are labeled "question" and others "discussion". If the link doesn't work, just copy and paste into your browser.
Hope that helps!
I would say is the butcher. The cold tempersture does keep its properties, what I don't know if you should sink the piece?
Melissa Bemrose thank you. I ordered a stomach which was cut open, rinsed and on ice. When I got it, i laid it in cold krebs solution. And I got much higher RNA concentrations now. Thanks! :-)
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