Concentrating DNA

From Protists

Background

• This is from edited from the end of the MoBio UltraClean GelSpin DNA Extraction Kit. To see the original directions, click on the link above and click on one of the 12400 word document links under the Protocols section of the page.
• This should be done if your DNA is too dilute for your purposes.

Protocol

1. Start with either your single tube of DNA or add several tubes of the same product into one tube.
2. Add 2uL of 5M NaCl.
3. Mix tube by inversion several times.
4. Add 100uL of 100% Ethanol (plastic Ethanol bottle with molecular sieves in the bottom; stored in the ~4C walk-in).
5. Mix tube by inversion several times.
6. Centrifuge at max speed (16,400 rpm) for 5min.
7. Invert all liquid into MoBio DNA Extraction Kit Waste Bottle kept under the hood.
8. Dry residual ethanol with tube lid open in a speed vac or dessicator (covered with foil) or ambient air (inverted on clean kimwipe.
9. Resuspend precipitated DNA in desired volume of HPLC H2O or another acceptable alternative, such as a TE buffer. (Let sit for ~1-2hrs to redissolve DNA completely while flicking tube intermittently.)