If you fix adequately (no NBF) you can go with 1 René J. From: Mikael Niku <mikael.n...@helsinki.fi> To: histonet@lists.utsouthwestern.edu Sent: Wednesday, February 13, 2013 8:15 AM Subject: [Histonet] Tissue processing for laser microdissection and RNA isolation?
Hello! May I ask for your recommendations for tissue processing methods for laser microdissection and subsequent RNA isolation? I can think of at least the following protocols, each with significant drawbacks (and questions): 1) Traditional FFPE sections: + easy handling + RNA is safe (but see below) + good morphology - RNA is fixed too, so yields are low and only small fragments retrieved Here, I'm pretty happy with Qiagen RNEasy FFPE kit - any other suggestions, maybe cheaper? 2) Traditional cryosections: + fairly good morphology + good yields, good quality RNA if everything goes well - RNA is easily destroyed - difficult to handle small samples without melting & destroying RNA I haven't been very succesful with this option. 3) RNALater -> cryosections: + RNA is safe + good RNA yields, good quality RNA - poor morphology - difficult to section We have problems making the tissues actually freeze for good sectioning - any tricks or tips here? 4) RNALater -> paraffin sections? I haven't tried this yet, but should be doable through ethanol etc. Found some references claiming that the RNA quality is poor. 5) New commercial innovations like Qiagen/Prenalytix Paxgene Tissue kit, claiming to achieve both RNA stabilization and good morphology. I haven't tried any of these yet. Pricing is the obvious drawback. With best regards, Mikael Niku, PhD Department of Veterinary Biosciences University of Helsinki _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
