Yu:quick miniprep

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(solutions need to be put into a table)
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Revision as of 16:05, 15 September 2009

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Contents

Procedure

  1. Spin down 1-3 mL culture
    1. 4000 RPM for 5min at 4°C
  2. Transfer pellet to eppendorf tube if not already in one
  3. Resuspend pellet in 200 μL P1
  4. Add 200 μL of P2
    1. Mix by inversion - solution will become very viscous
  5. Add 200 μL of P3
    1. Mix by inversion - white precipitate will form
  6. Spin mixture for 10 min at top speed
  7. Remove pellet with a sterile toothpick
  8. Add 600 μL Isopropanol
  9. Spin mixture for 3 min at top speed
    1. Decant supernatant
  10. Add 500 µl 70% EtOH and vortex briefly
  11. Spin mixture for 3 min at top speed
    1. Decant supernatant
  12. Air dry pellet
  13. Resuspend pellet if 50-100 μL TE8

Solutions

P1

       50 mM Tris pH8.0 2.5 ml – 1.0M

20 mM EDTA 2.0 ml – 0.5M H20 to 50 ml

P2

	0.2N NaOH	2.0 ml – 5.0N

1% SDS 2.5 ml – 20% H20 to 50 ml

P3

3M NaOAc pH4.8

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