Plasmid DNA Mini-prep for Fluorescent DNA Sequencing
    (This protocol was originally obtained from Diane England and modified by Andy Blasband, ABI June 15, 1991.)

    1. Pellet 1.5ml of culture grown in terrific broth (recommended) for 1 minute in a microcentrifuge. Remove all the supernatant by aspiration. If larger quantities of DNA are desired then spin down 3ml of culture.
    2. Resuspend pellet in 200ul of GTE buffer (50mM Glucose, 25mM Tris pH 8.0, 10mM EDTA pH 8.0) by pipetting up and down.
    3. Add 300ul of 0.2N NaOH/1% SDS buffer (freshly made), mix by tube inversion. Do Not Vortex . Incubate on ice for 5 minutes.
    4. Add 300ul of 3.0M KOAc, pH 4.8 (3.0M NaOAc pH 4.8 also works). Mix by tube inversion. Do Not Vortex . Incubate on ice for 5 minutes.
    5. Spin in microcentrifuge for 10 minutes at room temperature and transfer supernatant (approximately 700 to 750ul) to a new tube.
    6. Add RNase A to a final concentration of 20ug/ml (approximately 2ul of 10 mg/ml stock). Incubate at 37C for 20 minutes.
    7. Do two chloroform extractions using 400ul/extraction. (The tube capacity is too small do do equal volume extractions, half volume works fine.) Do not use phenol, use straight chloroform. Mix layers by shaking not vortexing. Spin extractions for 1 minute to separate phases and remove aqueous phase to a clean tube.
    8. Precipitate DNA by adding an equal volume of isopropanol (700 to 750ul). Place tube on ice for 10 minutes.
    9. Pellet DNA for 15 minutes at room temperature. Rinse pellet with 500ul of 70% ETOH. Pour off ETOH and dry the pellet in a speed vac.
    10. Dissolve pellet in 32ul of dH2O. Add 8ul of 4M NaCl and 40ul of 13% PEG8000. Mix well and on ice for 20 minutes, longer incubations will improve yields.
    11. Spin for 15 minutes at 4C. Remove supernatant (be careful as the pellet is translucent at this point) and rinse pellet with 500ul of 70% ETOH.
    12. Pour off ETOH, dry pellet in a speed vac and resuspend in 20 to 30ul of water.

    Terrific Broth
    Add 100ml of a sterile solution of 0.17M KH2P04 and 0.72M K2HPO4 to 900ml of base broth. (Base broth = 12g of Bacto-tryptone, 24g of Bacto-yeast extract, 4ml of glycerol, q.v. to 900ml with dH2O and autoclave).