I am currently performing RNA extractions from FACS sorted nuclei. The yields have been good. However, purity measures (i.e. 260/280, 260/230 ratios) have been poor. I suspect phenol contamination or something of the sort. I am considering including extra washing steps/other protocol alterations. Does anybody have any suggestions for this?
Many thanks,
Rob.
hi
1- you can reduce amount of zymo nuclei for better lysis , ...
2- test" RNeasy Mini Kit" from qiagen company .
3- Follow the 2 steps of washing the column well.
good luck
I can agree with Reza Hadavi on the use of the "Qiagen RNeasy Mini Kit". We've used that kit for several years and had decent results using it. Another suggestion would be upgrading to automatic extraction with magnetic beads (tho this will be expensive). We've had excellent results with magnetic bead extractions of RNA. Good luck with your experiment.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA