ABSTRACT
EDTA (ethylenediaminetetraacetic acid), pH 8 4. TE buffer: 10 mM Tris, 1 mM EDTA, pH 7.5 5. Lysis buffer: 1% sodium dodecyl sulfate (SDS) in 0.2 M
NaOH 6. Potassium acetate (5 M, pH 4.8) 7. RNase A (in the freezer) 8. Isopropanol (kept cold) 9. Ammonium acetate (7.5 M) 10. PCIA (phenol/ chloroform/ isoamyl alcohol) reagent 11. Isopropanol (kept cold) 12. Ammonium acetate (7.5 M) 13. DNA-grade absolute ethanol 14. Restriction enzymes and buffers 15. Electrophoresis buffer (tris-acetate-EDTA [TAE]), 10X
concentration 16. Agarose 17. Ethidium bromide 18. Gel tray and comb; lab tape 19. Gel loading buffer “6X” (50% v/ v glycerol and 0.5% w/ v
bromophenol blue) 20. DNA ladder(s)
1. For each miniprep, pipette 1.5 mL of the bacterial culture into a microcentrifuge tube. Spin the culture at top speed in a tabletop microcentrifuge for 60-90 seconds. Pipette off or dump out the supernatant, making sure no excess medium remains.
