Griffitts:Miniprep

Standard Qiagen Miniprep

Materials

 * Qiaprep Spin Kit (27106)
 * TE buffer
 * 1.5 mL microcentrifuge tubes (1 per culture)

Procedure

 * Grow DH5α with appropriate plasmid in 4 mL liquid LB+antibiotic for 12–18 hours
 * Label your spin columns and microcentrifuge tubes
 * Centrifuge 1.6 mL of each culture in a microcentrifuge tube at full speed for 30 seconds
 * Dump and tap out supernatant
 * Resuspend pellet in 250 μL P1
 * The P1 is kept in the fridge
 * Make sure to use P1 that has RNAse added
 * Add 250 μL P2
 * Immediately invert 3 times
 * Let stand for 2 minutes
 * Add 350 μL N3
 * Immediately invert 8 times
 * Centrifuge for 6 minutes at full speed
 * Carefully move the supernatant (~820 μL) to a spin column
 * Centrifuge for 30 seconds at full speed
 * Remove flowthrough
 * Add 700 μL of wash buffer PE to column
 * Centrifuge for 30 seconds at full speed
 * Remove flowthrough
 * Centrifuge for another 60 seconds at full speed
 * Move column to a labeled 1.5 mL microcentrifuge tube
 * Add 50–100 μL TE directly to disk
 * Don't touch the membrane with your tip
 * Let stand 1–2 minutes
 * Centrifuge for 60 seconds at full speed to elute DNA

Adapted from Qiagen handbook