Promega Maxi Prep (without vacuum manifold)
- Pellet 100-500ml cell solution in a centrifuge bottle at 4,986 rpm (5,000g) for 10 min.
- Discard supernatant, Resuspend in 15ml Cell Resuspension Solution
- Add 15ml Cell Lysis Solution, invert up to 20 times
- Add 15ml Neutralization Solution, invert several times
- Spin at 8,344rpm (14,000g) for 15min
- Run supernatant through filter paper into a graduated cylinder
- Transfer to a new centrifuge bottle and add 50% volume isopropanol
- Spin at 8,344rpm (14,000g) for 15min
- Discard supernatant, resuspend in 2ml TE
- Add 10ml DNA Purification Resin, swirl
- Pour into shredder column, let drain
- Spin at 2,500rpm (1,300g) with a swinging basket rotor for 5min, pour off
- Add 25ml Column Wash Solution
- Spin at 2,500rpm (1,300g) with a swinging basket rotor for 5min
- Add 5ml 80% ethanol, let drain
- Spin at 2,500rpm (1,300g) with a swinging basket rotor for 5min, pour off
- Spin again, discard collection tube
- Place column in a new 50ml tube and add 1.5ml 65° water, wait 1 min
- Spin at 2,500rpm (1,300g) with a swinging basket rotor for 5min
- Pour into syringe/filter set up and filter into a 1.5ml tube
- Spin at 8,344rpm (14,000g) for 1 min
- Transfer supernatant to a new tube.