Gene Clean Protocol

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14 mL of NEW concentrate + 280 mL of DI H2O à mix

Add 310 mL 100 % EtOH à mix

Store NEW Wash at -20C

1. Add 3 volumes of NaI stock to band (~250 + 750 uL NaI). Incubate for 5 min. at 45 – 55C to melt agarose.

2. Vortex Glassmilk mix until there is an even suspension. May take a minute or so – hold tube horizontally. 5 uL of Glassmilk to 5 ug of DNA.

3. Add 5 uL of Glassmilk to DNA / NaI solution. Mix and place on ice for 5 min. Mix every 1 – 2 min. to keep Glassmilk in suspension. If volume is > 1.5 mL allow 15 min. binding time.

4. Pellet reaction – quick spin. Remove supernatant. Respin. Remove remaining supernatant. Wash pellet 1 X with NaI stock (200 – 400 uL). Spin. Remove supernatant.

5. Wash pellet 3 X with NEW Wash:

Add 10 – 50 volumes of NEW Wash (200 – 700 uL). Resuspend with pipet tip. Spin. Remover supernatant. Repeat 2 X.

6. Elute DNA from glassmilk beads.

Resuspend pellet in TE buffer (amount = amount (uL) of Glassmilk added in step 3. Incubate at 45 – 55C for 2 – 3 min. Spin 30 seconds. Remover supernatant + DNA to fresh tube. Repeat elution 1 X.

7. Spin supernatants 1 X again. Remove to fresh tube. 10 uL à 2 ug.