Protocol/4

(Difference between revisions)

Revision as of 01:05, 26 October 2010

Protocol 4: Plasmid Miniprep

Reagents:

Procedure:

Transfer 1.5ml of culture to Eppendorf.
Spin down at 13 000 rpm.
Discard supernatant and follow procedure on p. 22 of [http://www.qiagen.com/literature/render.aspx?id=370 QIAprep Miniprep Handbook].
Determine concentration of purified plasmid with nanodrop.

Notes:

Low speed pellet is easier to resuspend.
To resuspend, take an empty micro centrifuge rack and rub the tube quickly over the apparatus and vortex.
Step 2 in Qiagen, the alkaline lysis with buffer P2, should be brief to avoid damaging DNA.

@@ Line 14: / Line 14: @@
-What you will need:
+Reagents:
 * Overnight culture
 * Qiagen plasmid miniprep kit
 Procedure:
-* Transfer 1.5 ml of culture to Eppendorf.
+* Transfer 1.5ml of culture to Eppendorf.
 * Spin down at 13 000 rpm.
 * Discard supernatant and follow procedure on p. 22 of [http://www.qiagen.com/literature/render.aspx?id=370 QIAprep Miniprep Handbook].
-* Determine concentration of purified plasmid with nanodrop
+* Determine concentration of purified plasmid with nanodrop.
-==Notes==
+Notes:
 * Low speed pellet is easier to resuspend.
-* To resuspend, take an empty micro centrifuge rack and rub the tube quickly over the holes, so it makes a "quacking" sound. It's pretty fun and works pretty good. Vortex after too.
+* To resuspend, take an empty micro centrifuge rack and rub the tube quickly over the apparatus and vortex.
 * Step 2 in Qiagen, the alkaline lysis with buffer P2, should be brief to avoid damaging DNA.
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