Yu:quick miniprep: Difference between revisions
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(solutions need to be put into a table) |
m (fixed solution format) |
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====P1==== | ====P1==== | ||
{| border="1" cellpadding="10" cellspacing="0" | |||
|width="300px" valign="top"|'''Final Concentration''' | |||
|width="100px" valign="top"|'''For 50 mL''' | |||
|width="100px" valign="top"|'''For 100 mL''' | |||
|- | |||
|50 mM Tris pH8.0 | |||
|2.5 mL of 1M Tris pH8 | |||
|5 mL of 1M Tris pH8 | |||
|- | |||
|20 mM EDTA | |||
|2.0 mL 0f 0.5 M EDTA | |||
|4.0 mL 0f 0.5 M EDTA | |||
|- | |||
|milliQ H<sub>2</sub>O | |||
|to 50 mL | |||
|to 100 mL | |||
|} | |||
====P2==== | ====P2==== | ||
{| border="1" cellpadding="10" cellspacing="0" | |||
|width="300px" valign="top"|'''Final Concentration''' | |||
|width="100px" valign="top"|'''For 50 mL''' | |||
|width="100px" valign="top"|'''For 100 mL''' | |||
|- | |||
|0.2N NaOH | |||
|1 mL of 10 N NaOH | |||
|2 mL of 10 N NaOH | |||
|- | |||
|1% SDS | |||
|2.5 mL 0f 20% SDS | |||
|5 mL 0f 20% SDS | |||
|- | |||
|milliQ H<sub>2</sub>O | |||
|to 50 mL | |||
|to 100 mL | |||
|} | |||
====P3==== | ====P3==== | ||
3M NaOAc pH4.8 | 3M NaOAc pH4.8 |
Latest revision as of 13:33, 18 September 2009
Procedure
- Spin down 1-3 mL culture
- 4000 RPM for 5min at 4°C
- Transfer pellet to eppendorf tube if not already in one
- Resuspend pellet in 200 μL P1
- Add 200 μL of P2
- Mix by inversion - solution will become very viscous
- Add 200 μL of P3
- Mix by inversion - white precipitate will form
- Spin mixture for 10 min at top speed
- Remove pellet with a sterile toothpick
- Add 600 μL Isopropanol
- Spin mixture for 3 min at top speed
- Decant supernatant
- Add 500 µl 70% EtOH and vortex briefly
- Spin mixture for 3 min at top speed
- Decant supernatant
- Air dry pellet
- Resuspend pellet if 50-100 μL TE8
Solutions
P1
Final Concentration | For 50 mL | For 100 mL |
50 mM Tris pH8.0 | 2.5 mL of 1M Tris pH8 | 5 mL of 1M Tris pH8 |
20 mM EDTA | 2.0 mL 0f 0.5 M EDTA | 4.0 mL 0f 0.5 M EDTA |
milliQ H2O | to 50 mL | to 100 mL |
P2
Final Concentration | For 50 mL | For 100 mL |
0.2N NaOH | 1 mL of 10 N NaOH | 2 mL of 10 N NaOH |
1% SDS | 2.5 mL 0f 20% SDS | 5 mL 0f 20% SDS |
milliQ H2O | to 50 mL | to 100 mL |
P3
3M NaOAc pH4.8