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Protocol/4
From 2010.igem.org
(Difference between revisions)
| Line 14: | Line 14: | ||
| - | + | Reagents: | |
* Overnight culture | * Overnight culture | ||
* Qiagen plasmid miniprep kit | * Qiagen plasmid miniprep kit | ||
| + | |||
Procedure: | Procedure: | ||
| - | * Transfer 1. | + | * Transfer 1.5ml of culture to Eppendorf. |
* Spin down at 13 000 rpm. | * 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]. | * 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: | ||
* Low speed pellet is easier to resuspend. | * Low speed pellet is easier to resuspend. | ||
| - | * To resuspend, take an empty micro centrifuge rack and rub the tube quickly over the | + | * 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. | * Step 2 in Qiagen, the alkaline lysis with buffer P2, should be brief to avoid damaging DNA. | ||
| + | |||
[[Team:Alberta/Notebook/protocols| Back]] | [[Team:Alberta/Notebook/protocols| Back]] | ||
{{Team:Alberta/endMainContent}} | {{Team:Alberta/endMainContent}} | ||
Revision as of 01:05, 26 October 2010
Protocol 4: Plasmid Miniprep
Reagents:
- Overnight culture
- Qiagen plasmid miniprep kit
Procedure:
- Transfer 1.5ml of culture to Eppendorf.
- Spin down at 13 000 rpm.
- Discard supernatant and follow procedure on p. 22 of 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.
