Hi fellow sciency friends,
I have a protocol for IF-FISH that says to use RNaseOut (Invitrogen) in my block at 1:100. I'm doing the IF-FISH on coverslips in a 6-well plate and I'm using 2mL block for blocking and 2mL block for primary (could potentially go down on this volume).
RNaseOut is supplied as 5000U (40U/uL) in 125uL. This means that I would have to use a whole tube to only get 12.5mL of block which is, frankly, ridiculously expensive to me (@ ~$180/tube).
Does anyone else use RNaseOut and if so, is this an accurate dilution to use? Invitrogen website says 40U/20uL which would be even more concentrated of a dilution. Alternatively, does anyone use a different RNase inhibitor or have any other insight into blocking RNA degradation during IF (necessity, perhaps?). In the words of Leeloo Dallas Multipass, "PLEAZ HALP". Thanks so much!
Mandy
Benjamin THOHA Thomas
I spoke with the individual that had initially taught me FISH (but just described the IF protocol as we weren't practicing that at the time)-- he said that the RNaseOut concentration is correct and that, instead of filling the well with block and primary to pipet a small volume of the solution directly onto the coverslip and cover with the non-exposed side of parafilm to create a parafilm-block-coverslip sandwich and significantly reduce the volume necessary.
- Badges
- Science topic