Hi,
I have been trying to grow primary myofibroblasts from UUO-injured mice kidneys. I used protocol, and grew cells for a duration of 14 days, cells grew well with 80-90% confluency at D14, and my plan was to do qPCR to determine enrichment of the myofibroblasts. However when I did the RNA extraction ( RN-easy microkit ( Qiagen) - I had extremely low yield ( close to none). Anyone has any input on how to troubleshoot this? It is definitely very frustrating to reach this step. Appreciate all your advise and comments
Thank you!
Hello Amirtha,
I am writing because I have a very similar problem with low RNA yields from primary cells, but from tumor origin. I have plenty of cells (~million) and visibly see the pellet prior to lysis of the cells using the Qiagen RNeasy Plus Mini kit. I do not see any responses here but I was wondering if you had any luck improving your yields? Any suggestions are much appreciated. Thank you in advance.
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