Homogenize pancreas for RNA extraction
Posted 21 February 2013 - 03:02 AM
I am wondering, which is the best method to homogenize the pancreas of a mice to extract the RNA?
Should i use the whole pancreas or weigh it ~ 30mg per each reaction as suggested by the RNeasy Mini kit?
Thank you very much.
Posted 22 February 2013 - 06:51 AM
Posted 22 February 2013 - 08:15 AM
I recommend you read a couple of technique papers on pancreas isolation. Due to the amount of RNAses present in the pancreas, isolating intact RNA from the pancreas is extremely difficult. In our lab, we do a quick resection (under 10 seconds) and snap freeze the tissue. We then pulvorize the tissue while frozen and hit with TRIZOL immediately.
There are other techinques that you can find on the web. You will have to find what works best for your lab setting. Just make sure to do Agilent for your RNA and calculate RIN to make sure you don't have degraded RNA.
Posted 23 February 2013 - 11:46 AM
Thanks alot guys for sharing your experiences and tips&tricks.
Have a nice day