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==Size selected DNA recovery from TBU-PAGE== *Prepare gel shearing assemblies by placing a 0.5mL tube (punched a hole in bottom using G22 needle) in a 1.5-mL tube. *Transfer the sliced gel in 0.5mL tubes. Centrifuge at 14000 rpm at RT for 3min. *After centrifuge, remove the 0.5-mL tubes. Add 500uL 1X TE with the sheared gel in 1.5-mL tubes. Transfer the mixture to a fresh 1.5-mL tube. *Incubate the gel-TE mixture at 37C for 1hr on the shaker (shaking at lowest speed) *Centrifuge the gel-TE buffer mixture at 14000 rpm at RT for 1.5min. The gel debris will be pellet. Transfer the supernatant into a Nanosep column filter. *Centrifuge the Nanosep columns at 14000 rpm at RT for 1.5min. The tiny gel debris will remain on the filter. Collect the flow through. *Perform DNA purification with EtOH/NaOAc method. **Sample + 2.5V 100% EtOH + 0.1V 3M NaOAc + 1.5uL Glycoblue. **Gentaly invert the tube 20times. Incubate at -80C for 25min (or longer) **Centrifuge at 4C, 10,000xg for 25min. Remove the supernatant(pour + 20p tips). Wash the pellet with 75% EtOH. Gentaly invert the tube 20times. **Centrifuge at 4C, 10,000xg for 5min. Remove the supernatant(pour + 20p tips). Air dry the pellet in PCR hood (~10min). **Elute the DNA in 20uL H2O.
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