Miniprep/TENS miniprep

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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<br>
+
: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
+
:5 mL 1M Tris-HCl pH 8.0
-
*12.5 mL 20% SDS
+
:12.5 mL 20% SDS
-
*5 mL 10N NaOH
+
:5 mL 10N NaOH
-
*Enough H20 to 500 mL
+
:Enough H20 to 500 mL
[[Category:Protocol]]
[[Category:Protocol]]

Revision as of 21:16, 15 May 2012

back to protocols

Contents

Background

TENS method of minipreps. Fast but dirty.

Protocol

  1. 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.
  2. Remove and discard the supernatent.
  3. Resuspend either in 50 μL of LB or P1 buffer or TE/RNAse
  4. Add 300 μL TENS buffer
  5. Mix by inverting 5 times.
  6. Add 100 μL 3M NaAc pH 5.2
  7. Mix again
  8. Spin down at top speed for 5 minutes
  9. Transfer supernatant to a new tube (pouring works)
  10. Add 1 mL 100% EtOH (ideally ice cold)
  11. Spin down at top speed for 5 minutes (ideally at 4˚C.)
  12. Wash with 0.5 mL of 70% EtOH
  13. Pour out the supernatant
  14. Spin again to find the rest of the supernatant
  15. Air dry for 10 minutes
  16. Resuspend the dried pellet in 30 μL of water or TE buffer.
  17. 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

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