Miniprep/TENS miniprep: Difference between revisions
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m (New page: {{back to protocols}} ==Background== TENS method of minipreps. Fast but dirty. ==Protocol== #Transfer 1.5 mL of an overnight culture containing your plasmid to an eppendorf tube and spin ...) |
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===TE=== | ===TE=== | ||
10 mM Tris-HCl, pH 8 | :10 mM Tris-HCl, pH 8 | ||
1 mM EDTA | :1 mM EDTA | ||
==TENS== | ===TENS=== | ||
(500 mL recipe) | (500 mL recipe) | ||
:5 mL 1M Tris-HCl pH 8.0 | |||
:12.5 mL 20% SDS | |||
:5 mL 10N NaOH | |||
:Enough H20 to 500 mL | |||
[[Category:Protocol]] | [[Category:Protocol]] | ||
Revision as of 18:16, 15 May 2012
| back to protocols | ||
Background
TENS method of minipreps. Fast but dirty.
Protocol
- Transfer 1.5 mL of an overnight culture containing your plasmid to an eppendorf tube and spin at 5000 rpm for 5 min in a tabletop centrifuge to pellet the cells.
- Remove and discard the supernatent.
- Resuspend either in 50 μL of LB or P1 buffer or TE/RNAse
- Add 300 μL TENS buffer
- Mix by inverting 5 times.
- Add 100 μL 3M NaAc pH 5.2
- Mix again
- Spin down at top speed for 5 minutes
- Transfer supernatant to a new tube (pouring works)
- Add 1 mL 100% EtOH (ideally ice cold)
- Spin down at top speed for 5 minutes (ideally at 4˚C.)
- Wash with 0.5 mL of 70% EtOH
- Pour out the supernatant
- Spin again to find the rest of the supernatant
- Air dry for 10 minutes
- Resuspend the dried pellet in 30 μL of water or TE buffer.
- Do your molecular bio
Buffers
TE
- 10 mM Tris-HCl, pH 8
- 1 mM EDTA
TENS
(500 mL recipe)
- 5 mL 1M Tris-HCl pH 8.0
- 12.5 mL 20% SDS
- 5 mL 10N NaOH
- Enough H20 to 500 mL
