Recipes/MethylCell

From Kimmel Lab Wiki

3% Methyl Cellulose

Quantity	Ingredients	Location
49.5 ml	Embryo medium	Area B, counter
0.5 ml	1M HEPES	Area M, door shelf – upper left
1.5 g	Methyl cellulose; cat# M0387 SIGMA	Area M, door shelf – 2nd row from upper right

Directions

1. Preheat EM and HEPES in a 50 ml culture tube in water bath to ~50 degrees C
2. Shake the methyl cellulose container before measuring to break up any lumps and homogenize the various components
3. Add methyl cellulose to 50 ml culture tube & shake until there are no lumps
4. Put back in the water bath for at least 30 minutes, longer is better. *Note* Solution will be cloudy.
5. Immerse in an ice bath, check and stir every 10 minutes
6. When the solution is a thick homogeneous gel with an even suspension of tiny bubbles making it opaque, refrigerate over night.
7. After bubbles disappear, aliquot into about 5 ml amounts and store at 4C.
8. Best used at your working temperature to prevent bubbling from a temperature change.