Could formaldehyde fixation and 70% ethanol permeabilization be carried out simultaneously, not sequentially during the sample treatment in RNA hybridization? what's the purpose of ethanol permeabilization since formaldehyde alone has fixed the cells ?
You can fix and permeabilize at the same time using ethanol/methanol + acidic acid (see the document below). I find that if I use ice-cold methanol without acidic acid, the nucleic acids will precipitate. For example, the DNA gets dragged out of the cells.
In many applications, Formaldehyde or PFA fixation is the least detrimental to the cells. Once you have the cells fixed, you can permeabilize with 70% ethanol without being too worried about distorting biological structures.
This document might come in handy:
http://www.abcam.com/ps/pdf/protocols/fixation_permeabilization.pdf
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA