RNAlater recipe

Submerge all your equipment and glassware in 1% bleach to get rid of nuclease

DEPC treated water

1. Add 1mL DEPC in 1L ddH2O (0.1% (v/v) ); MIX it well.

2. Let it sit for 24h 0m 0s at25°C

3. Autoclave it to deactivate DEPC

4. Store the DEPC treated water at4°C

0.5 M Disodium dihydrate EDTA :

1. Add 18.61g disodium dihydrate EDTA in 100mL ddH2O

2. Adjust pH to 8.0 with NaOH while stiring

1 M tri-Sodium Citrate Dihydrate :

Add 29.4g tri-Sodium Citrate Dihydrate in 100mL ddH2O, stir to dissolve

1 M sulfuric acid :

Add 5.33mL sulfuric acid in100mLddH2O, stir to dissolve

Make a mixture of:

40mL of 0.5 M EDTA + 25mL 1 M tri-Sodium Citrate + 700g Ammonium Sulfate +935mL ddH2O ; stir on a hot plate with low heat until ammonium sulfate dissolved

Allow it to cool; adjust the pH of the solution to pH 5.2 using 1 M sulfuric acid

The final concentration: 25 mM Sodium Citrate, 10 mM EDTA, 70 g ammonium sulfate in 100 ml ddH2O; pH 5.2.

Transfer to screw top, nuclease-free bottles; label the date of make; store at 4°C