Preparation of Denaturing Agarose Gel for RNA Analysis
Melt 1.0 g agarose in 87 ml of DEPC water , by dispersing the agarose uniformly and heating in a microwave until all particles are dissolved .
Bring the melted agarose to 60 °C .
Add 10 ml 10X MOPS Buffer and 3 ml 37 % formaldehyde .
Pour standard agarose gel gel .
USE A FUME HOOD !
FORMALDEHYDE IS VOLATILE AND TOXIC .
WORK IN A HOOD FROM THIS POINT FORWARD .
Allow gel to set for 1 hour .
Run gel in 1X MOPS Buffer .
Mix 40 µl sample buffer with 10 µl sample , heat to 55 °C 15 minutes .
Add 10µl of the following : 50 % glycerol 1 mM EDTA 0.3 % each bromophenol blue and 0.3 % xylene cyanol .
