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Protocols
Nucleic acid isolation
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Protocols

Brian Krueger, PhD
Columbia University Medical Center
Price Lab
DNA Ethanol Precipitation
Description: 2 methods for precipitating DNA after isolation. Ammonium acetate and sodium acetate methods.
Date Uploaded: Tue, Mar 25, 2008, 1:52 pm CDT

Brian Krueger, PhD
Columbia University Medical Center
Price Lab
RNA Extraction
Description: This is an extraction protocol using invitrogen's Trizol reagent
Date Uploaded: Tue, Mar 25, 2008, 2:20 pm CDT
Aklaline Lysis Plasmid Isolation (High Throughput)
Description: A quick and dirty protocol that yields double stranded plasmid template of approximately 3 mg per sample. Colonies are grown in 96 deep well plates (~1 mL sample volume), and then subjected to alkaline lysis.
Date Uploaded: Fri, Sep 03, 2010, 1:14 pm CDT
Bead Beating DNA Extraction Protocol
Description: Soils can be a pain in the rear to extract DNA from. We've tried to work this protocol to eliminate as much bias as possible from the final prep. Bead beat too long and you'll ruin the DNA from your Gram negatives. Bead beat too short and you risk not cracking open your Gram positives. To increase final DNA yields, you could also try adding 1 to 2 uL of GlycoBlue with your 95% EtOH precipitation.
Date Uploaded: Fri, Sep 03, 2010, 1:16 pm CDT
EtOH Precipitation
Description: The following PDF is an article that was published by the now defunct Bethesda Research Laboratories back in 1985 (before half the people who read this were even born). It discusses the effects of both incubation time and temperature, as well as centrifugation time. The moral is ... spin your DNA prep down for at least 30 minutes. **I don't think posting this poses any copyright issues. My apologies if it does.**
Date Uploaded: Fri, Sep 03, 2010, 1:07 pm CDT
Acetone/EtOH Bacterial RNA Isolation
Description: A protocol used in our lab for isolation of RNA from bacterial cultures. This protocol works equally well for Gram positive and Gram negative bacteria (we've done it with both). The acetone/ethanol mixture is great for long term storage of samples prior to RNA isolation. Cells are theoretically permeabilized and proteins (including RNAses) are inactivated.
Date Uploaded: Fri, Sep 03, 2010, 1:15 pm CDT
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