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Lithium Chloride RNA Cleanup Protocol

LiCl cleanup is used after Trizol RNA extraction protocols.

 

Procedure:

1)      Add 1:1 volume 4M LiCl and place on ice for 1 - 2 hours.

2)      Centrifuge at full speed for 10 minutes and remove supernatant.

3)      *Resuspend in 100uL TE or nuclease-free water.

4)      Precipitate RNA:

a.       Add 0.1 vol 3M NaOAc (10uL)

b.      Add 2.5 vol 95% EtOH (250uL)

5)      Spin for 10 minutes at full speed and remove the supernatant

6)      Air dry pellet

7)      Resuspend pellet in 20uL nuclease-free water.

Solution Recipes:

4M LiCl (42.39g/M)

            25.4g LiCl in 250mL nuclease-free water

 

1X TE Buffer

-1mL 1M Tris-Cl (6.06g Tris base + 40mL nuc. free water, pH to 8.0 with concentrated HCl, adjust total volume to 50mL)

-0.2mL .5M EDTA (Ambion)

-Adjust volume to 100mL

 

3M Sodium Acetate (NaOAc-3H20 is 136.08g/M)

            81.6g NaOAc-3H20  in 200mL nuclease free water