Team:Alberta/Notebook/protocols/miniprep

From 2010.igem.org

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==General Protocols==
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/invitro_biobyte_assembly | In Vitro BioByte Assembly]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/LB | LB Plates and Broth]]
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*[[Team:Alberta/Notebook/protocols/transformations |Transformations]]
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*[[Team:Alberta/Notebook/protocols/overnight |5mL Overnight ]]
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*[[Team:Alberta/Notebook/protocols/glycerol | Glycerol Stock ]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/miniprep | Plasmid Miniprep ]]
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*[[Team:Alberta/Notebook/protocols/digest | Restriction Digest ]]
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*[[Team:Alberta/Notebook/protocols/vector_dephos | Vector Dephosphorylation]]
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*[[Team:Alberta/Notebook/protocols/ligation | Ligation ]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/agarose_gel | Agarose Gel Electrophoresis ]]
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*[[Team:Alberta/Notebook/protocols/gel_extraction | Gel Extraction ]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/pcr | PCR]]
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*[[Team:Alberta/Notebook/protocols/colony_pcr | Colony PCR ]]
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*[[Team:Alberta/Notebook/protocols/pcr_purification | PCR Purification ]]
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-----------------------------
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*[[Team:Alberta/Notebook/protocols/labelling | Sample Labelling Conventions]]
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*[[Team:Alberta/Notebook/protocols/sequencing | Fluorescent Sequencing Reaction]]
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*[[Team:Alberta/Notebook/protocols/primer_design | Primer Design]]
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-----------------------------
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Plasmid Miniprep
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==Plasmid Miniprep==
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Reagents:
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===Reagents:===
* Overnight culture
* Overnight culture
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Procedure:
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===Procedure:===
* Transfer 1.5ml of culture to Eppendorf.
* Transfer 1.5ml of culture to Eppendorf.
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Notes:
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===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 apparatus and vortex.
* To resuspend, take an empty micro centrifuge rack and rub the tube quickly over the apparatus and vortex.

Latest revision as of 02:34, 27 October 2010

TEAM ALBERTA


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.