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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.