IGEM:Brown/2007/Lab Protocols/10min E coli Miniprep

Stocks TENS (for 100ml) *1ml 10N NaOH *1ml 20% SDS *1ml 1M Tris pH 7.5 *200 microliters 0.5 M EDTA *96.8 mL water 3 M NaOAc pH 5.2

Procudure


 * 1) Spin 1.5 ml o/n culture E coli in an eppendorf for 10 s
 * 2) Decant s/n and resuspend in the remaining volume
 * 3) Add 300 microliters TENS. Vortex to mix completely
 * 4) Add 150 microliters 3 M NaOAC. Mix by inversion
 * 5) Spin 10 min room temperature (debris, genomic DNA)
 * 6) Transfer s/n to fresh tube
 * 7) Add 900 microliters ice cold EtOH. Mix thoroughly
 * 8) Sping 15 min room temperature
 * 9) Wash pellet 1x 70% EtOH (optional)
 * 10) Dry pellet speed-vac
 * 11) Resuspend in 100 microliter TE. Use 1.5-3 microliters for digest
 * 12) Add 1 microliter1 1mg/ml RNAase to 10 microliter digest