This is a cached page for the URL (http://www.elabprotocols.com/viewer/?id=167). To see the most recent version of this page, please click here.
Protocol Online is not affiliated with the authors of this page nor responsible for its content.
About Cache
RNA gel electrophoresis - eLabJournal.com
Average Rating:
Rate protocol:

Category: Experimental Procedures
Author: Admin eLabJournal

Labels: Electrophoresis, RNA

Materials
 

MOPS buffer 10X
Agarose
Formaldehyde (37%)
MOPS buffer 1X
Formamide
Ethidium bromide (100 mg ml-1)
Denaturation mix
Loading buffer: Orange G or Xylene cyanol/Bromophenol blue
Gel casting system
Gel running system
UV-transilluminator

Experiment settings

Volume of gel
ml 
Agarose percentage
Step 1
formaldehyde-agarose gel of ml :
Step 2
Dissolve in the microwave and allow to cool to 50°C
Step 3

Perform all next steps in the fumehood

Step 4
Add of ml  of 37%-formaldehyde and add 1-2 µl of ethidiumbromide
Step 5
Swirl and pour the agarose-solution into the gel tray, place a comb with the appropriate number of wells and allow the gel to solidify
Step 6
Remove the comb, place the gel in a gel running system such that the gel is submerged in 1X MOPS buffer
Step 7
Denature µg  of RNA in µl  of denaturation mix for 15 min at 55°C.
Step 8
Mix the denatured RNA samples with µl  of loading buffer and load the samples on gel
Step 9
Run the gel at 40 mA for 45 min and visualize the RNA in a UV-transilluminator.
  • Make sure that all used materials are RNase free.
  • Use gloves and change them regularly.