I am trying to isolate RNA from rats hippocampus and cortex. Somehow, i am not able to get more than 10 ng/µl. Is that normal? I am using Qiagen kit and it works for all different types of mouse or human tissues but not the rat! any idea what should be changed?
Hello..there may be some problem in sample, how old the sample is? What are the storage conditions of sample? I will suggest to isolate new fresh sample (Hippocampus and Cortex) and isolate the RNA on the same day by manual method as well as Qiagen kit. Then we can check what is the exact problem?
Gud luck..
Difficult to answer. Is it neonate rats? Otherwise, my guess would be some problem with your centrifuge
I have done RNA extraction quite extensively using culture hippocampal neurons obtained from rat. ,The yield was huge. I was using TRIZOL reagent. From my experience one guess would be the dissociation phase. How do you dissociate CTX and Hip?
Try Standard Trizol Procedure (if you're not interested in proteins). It gives you quite a good yield (compared to some commercial kits), is straight-foward, quick and reliable. Good luck
Hi guys, i checked every single step you mentioned and it did not change the yield. Thats why i am asking here. I need some creative ideas.. i used Trizol. the centrifigue is good to go, the samples are stored correctly in -80C by snap freezing. Unfortunately, new samples will not be possible.
Try to pick the phases (very carefully and should be neat and clean) which you get after phase separation. Trizol is a wonderful method and gives very high yield from hippocampus. Try to do steps more quickly. Slow processing over each steps greatly affects the yield. Good luck
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