Team:Alberta/Notebook/protocols/overnight

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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Protocol 2: 5mL Overnight
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==5mL Overnights==
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Reagents:
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===Reagents:===
* 10mL culture tube Use 16mm x 160mm or 16mm x 125 mm
* 10mL culture tube Use 16mm x 160mm or 16mm x 125 mm
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Procedure:
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===Procedure:===
* Pipet 1.5-20uL of the appropriate antibiotic to the appropriate working concentration into culture tube.
* Pipet 1.5-20uL of the appropriate antibiotic to the appropriate working concentration into culture tube.
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Notes:
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===Notes:===
For I0500, culture overnight; then add 1mM ITPG in morning and continue culture for another 3 hours. Then miniprep.
For I0500, culture overnight; then add 1mM ITPG in morning and continue culture for another 3 hours. Then miniprep.
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Latest revision as of 02:21, 27 October 2010

TEAM ALBERTA


5mL Overnights

Reagents:

  • 10mL culture tube Use 16mm x 160mm or 16mm x 125 mm
  • 5mL LB
  • Concentrated antibiotics
  • Single colonies on a plate (Best not to start O/N from glycerol stocks)


Procedure:

  • Pipet 1.5-20uL of the appropriate antibiotic to the appropriate working concentration into culture tube.
  • Add 5mL non-contaminated LB. Then add antibiotic.
  • Select single colony using a sterile instrument that has been cooled and inoculate the liquid culture.
  • Place culture tube in incubator at 37oC overnight shaking vigorously (250 rpm).


Notes:

For I0500, culture overnight; then add 1mM ITPG in morning and continue culture for another 3 hours. Then miniprep.


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