RNA pellet gone in the wrong concentration of ethanolRNA Trizol Re
Posted 06 November 2012 - 08:50 AM
I wanted to wash the pellet with 75% ethanol, but unfortunately, the concentration of the ethanol I prepared is 25%.
Then I votex the RNA pellet in 25% ethanol and did the centrifugation.I found that the pellet has gone away. And I add more ethanol high up to the right concentration. After that I did the centrifugation another time.
But still, I can not spin the RNA down to the bottom. Should I add NaCl or other salt to solve this problem? Do I need to incubate it in lower temperture? Is the carrier needed this time?
Really need some help.
Posted 06 November 2012 - 11:45 AM
Posted 06 November 2012 - 06:12 PM
in RNAzol you should add 0.4 ml 75% ethanol to 1ml of supernatant, which the final concentation of ethanol is 21%.
(400 ul x 75%) + (1000 ul x 0%) = 1400 ul x X% => X= 21.42 %
So something else is wrong.
One of them:
If the starting cells are less than 1Million, the RNA pellete is very tiny. and specially when RNA is clean, it is very dificult to see it in vial.
I have not used but you can use glycoblue or something else.
Edited by memari, 06 November 2012 - 06:22 PM.
Posted 07 November 2012 - 04:36 AM
Do you have the protocol which I should follow? I don't know how much salt should add in to the solution
Edited by cckishi, 07 November 2012 - 04:40 AM.
Posted 07 November 2012 - 05:51 AM