Team:Michigan/Protocols

From 2010.igem.org

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'''Cell Culture'''
'''Cell Culture'''
-
[[Media:6-28-2010_Protocol_for_making_cultures_from_a_-80C_freezer_stock.pdf|Making cultures from a -80C freezer stock]]
+
[[Media:6-28-2010_Protocol_for_making_cultures_from_a_-80C_freezer_stock.pdf|Making cultures from a -80°C freezer stock]]
[[Media:6-29-2010_Making_Frozen_Stoks.pdf|Making frozen stocks]]
[[Media:6-29-2010_Making_Frozen_Stoks.pdf|Making frozen stocks]]
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[[Media:OD600 Cell Dilution Protocol.pdf‎|OD600 Cell Dilution Protocol]]
[[Media:OD600 Cell Dilution Protocol.pdf‎|OD600 Cell Dilution Protocol]]
 +
 +
[[Media:Quick Transformation of E. coli Competant Cell Preperation.pdf| Competent Cell Preparation for Quick Transformation of E. coli]]
'''DNA Manipulation'''
'''DNA Manipulation'''
-
[[Media:Protocol_Heat_Shock_Transformation_2.pdf|Transformation-heat shock]]
+
[[Media:Direct Plating Transformation Protocol.pdf|Direct Plating Transformation Protocol]]
 +
*Very simple protocol
 +
*Twice as efficient as standard heat shock
 +
 
 +
[[Media:Protocol_Heat_Shock_Transformation_2.pdf|Transformation - heat shock]]
*Getting parts from the 360 well registry plates
*Getting parts from the 360 well registry plates
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*Heat shock
*Heat shock
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[[Media:General_Transformation_Protocol.pdf|Transformation-electroporation]]
+
[[Media:General_Transformation_Protocol.pdf|Transformation - electroporation]]
*Getting parts from the 360 well registry plates
*Getting parts from the 360 well registry plates
*Competent cell preparation
*Competent cell preparation
*Electroporation
*Electroporation
 +
 +
[[Media:Electroporation Protocol 2.pdf|Electroporation Protocol 2]]
 +
*Adopted from Jeremy Minty's protocol
 +
*General protocol with specific instructions for ERB/Lin lab equipment
 +
 +
[[Media:PCR Screening Protocol.pdf|PCR Screening Protocol]]
 +
*Efficient protocol for screening transformants after a ligation
[[Media:MODIFIED_Miniprep_Protocol.pdf|MODIFIED Miniprep]]
[[Media:MODIFIED_Miniprep_Protocol.pdf|MODIFIED Miniprep]]
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[[Media:General_DNA_Digest_Protocol.pdf|DNA Digest]]
[[Media:General_DNA_Digest_Protocol.pdf|DNA Digest]]
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[[Media:Protocol_for_Running_a_Gel.pdf|Running a Gel]]
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[[Media:_Digest_reagents_calculator.xls|Digest Reagents Calculator]]
 +
 
 +
[[Media:Protocol_for_Running_a_Gel.pdf|Gel Electrophoresis]]
[[Media:Protocol_for_DNA_Purification.pdf|DNA Purification]]
[[Media:Protocol_for_DNA_Purification.pdf|DNA Purification]]
 +
 +
[[Media:Quick_Ligation_Protocol.pdf|Quick Ligation Protocol]]
[[Media:8-8-2010_Protocol_for_gel_extraction.pdf|Gel extraction]]
[[Media:8-8-2010_Protocol_for_gel_extraction.pdf|Gel extraction]]
[[Media:8-10-2010_Ligation_Calculations_updated.pdf|Updated T4 DNA Ligase Protocol (Not quick ligase)]]
[[Media:8-10-2010_Ligation_Calculations_updated.pdf|Updated T4 DNA Ligase Protocol (Not quick ligase)]]
 +
*For additional information/questions, refer to [http://www.neb.com/nebecomm/tech_reference/modifying_enzymes/ligation_tips.asp NEB Ligation Tips]
*[[Media:Ligation_Protocol.pdf|Ligation]]
*[[Media:Ligation_Protocol.pdf|Ligation]]
**Ginko Bioworks Protocol
**Ginko Bioworks Protocol
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[[Media:8-6-2010_Biofilm_Formation_Experiment.pdf|Revised biofilm assay protocol]]
[[Media:8-6-2010_Biofilm_Formation_Experiment.pdf|Revised biofilm assay protocol]]
-
*[[Media:7-28-2010_Biofilm_Formation_Experiment.pdf|biofilm assay protocol]]
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*[[Media:7-28-2010_Biofilm_Formation_Experiment.pdf|Biofilm assay protocol]]
 +
 
 +
[[Media:Static+biofilm+quantification.pdf|Alex's biofilm assay protocol]]
 +
 
[[Media:7-31-2010_Flu_operon_primers.pdf|General primer design]]
[[Media:7-31-2010_Flu_operon_primers.pdf|General primer design]]
[[Media:7-31-2010_Ag43_flu_gene_WITH_MODIFIED_PRIMER_SITES.pdf|Detailed primer design]]
[[Media:7-31-2010_Ag43_flu_gene_WITH_MODIFIED_PRIMER_SITES.pdf|Detailed primer design]]
-
[[Media:8-4-2010_Colony_PCR_flu.pdf|colony PCR protocol]]
+
 
 +
[[Media:8-4-2010_Colony_PCR_flu.pdf|Colony PCR protocol]]
 +
 
 +
[[Media:NanR_Cloning.pdf|NanR cloning]]
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*Updated Colony PCR Protocol
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 +
[[Media:NA Extraction.pdf|NA Extraction Protocol]]
====Pili====
====Pili====
[[Media:FimB_Ligation_Protocol.pdf|FimB Ligation]]
[[Media:FimB_Ligation_Protocol.pdf|FimB Ligation]]
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 +
[[Media:FimB_PCR.pdf|FimB PCR]]
 +
 +
[[Media:Flocculation_protocol.pdf|Flocculation Protocol]]
====Quorum Sensing====
====Quorum Sensing====

Latest revision as of 00:01, 27 October 2010


Michigan Header




Protocols

Using Lab Equipment

Obtaining Deionized Water in the ERB

ERB Spectrophotometer

Enigeering Research Building Autoclave

Epifluorescence Microscope Usage - H.H. Dow

Cell Culture

Making cultures from a -80°C freezer stock

Making frozen stocks

P. putida KT2440 antibotic resistance tolerance

Culturing CGSC Strains

OD600 Cell Dilution Protocol

Competent Cell Preparation for Quick Transformation of E. coli

DNA Manipulation

Direct Plating Transformation Protocol

  • Very simple protocol
  • Twice as efficient as standard heat shock

Transformation - heat shock

  • Getting parts from the 360 well registry plates
  • Competent cell preparation
  • Heat shock

Transformation - electroporation

  • Getting parts from the 360 well registry plates
  • Competent cell preparation
  • Electroporation

Electroporation Protocol 2

  • Adopted from Jeremy Minty's protocol
  • General protocol with specific instructions for ERB/Lin lab equipment

PCR Screening Protocol

  • Efficient protocol for screening transformants after a ligation

MODIFIED Miniprep

Nanodrop

DNA Digest

Digest Reagents Calculator

Gel Electrophoresis

DNA Purification

Quick Ligation Protocol

Gel extraction

Updated T4 DNA Ligase Protocol (Not quick ligase)

  • For additional information/questions, refer to [http://www.neb.com/nebecomm/tech_reference/modifying_enzymes/ligation_tips.asp NEB Ligation Tips]
  • Ligation
    • Ginko Bioworks Protocol
    • Jeremy Minty Protocol

Group-Specific Protocols

Oil Sands

Revised biofilm assay protocol

Alex's biofilm assay protocol

General primer design

Detailed primer design

Colony PCR protocol

NanR cloning

  • Updated Colony PCR Protocol

NA Extraction Protocol

Pili

FimB Ligation

FimB PCR

Flocculation Protocol

Quorum Sensing

Logic Table

Lsr Circuit Test Protocols

General Outline for Biobrick Manipulations

Igem order of procedures.jpg