Team:Washington/Protocols

From 2011.igem.org

(Difference between revisions)
m
 
(40 intermediate revisions not shown)
Line 1: Line 1:
{{Template:Team:Washington/Templates/Top}}
{{Template:Team:Washington/Templates/Top}}
__NOTOC__
__NOTOC__
 +
 +
<center><big><big><big><big>'''Protocols'''</big></big></big></big></center><br><br>
=General Protocols=
=General Protocols=
 +
 +
[https://2011.igem.org/Team:Washington/Protocols/gel_electrophoresis  General Agarose Gel Electrophoresis]
[https://2011.igem.org/Team:Washington/Protocols/PCR  General PCR Protocol]
[https://2011.igem.org/Team:Washington/Protocols/PCR  General PCR Protocol]
Line 16: Line 20:
[https://2011.igem.org/Team:Washington/Protocols/Competent  Competent Cell Prep Protocol]
[https://2011.igem.org/Team:Washington/Protocols/Competent  Competent Cell Prep Protocol]
-
[https://2011.igem.org/Team:Washington/Protocols/Kunkel Kunkel Mutagensis]
+
[https://2011.igem.org/Team:Washington/Protocols/Kunkel Kunkel Mutagenesis]
-
[http://www.bio.davidson.edu/Courses/molbio/kunkel/kunkel.html Overview of how Kunkel Mutagensis works]
+
[http://www.bio.davidson.edu/Courses/molbio/kunkel/kunkel.html Overview of the Kunkel Mutagenesis process]
[https://2011.igem.org/Team:Washington/Protocols/expression_purification Standard 1L Expression Purification]
[https://2011.igem.org/Team:Washington/Protocols/expression_purification Standard 1L Expression Purification]
Line 27: Line 31:
[https://2011.igem.org/Team:Washington/Protocols/CompDesign  Computational Protein Design]
[https://2011.igem.org/Team:Washington/Protocols/CompDesign  Computational Protein Design]
-
 
-
[https://2011.igem.org/Team:Washington/Protocols/gel_electrophoresis  Gel Electrophoresis]
 
[https://2011.igem.org/Team:Washington/Protocols/Glycerol_Stocks  Glycerol Stocks]
[https://2011.igem.org/Team:Washington/Protocols/Glycerol_Stocks  Glycerol Stocks]
Line 36: Line 38:
[https://2011.igem.org/Team:Washington/alkanebiosynthesis Alkane Biosynthesis media and extraction]
[https://2011.igem.org/Team:Washington/alkanebiosynthesis Alkane Biosynthesis media and extraction]
 +
 +
[https://2011.igem.org/Team:Washington/alkanebiosynthesis_cloning Alkane Biosynthesis cloning]
 +
 +
[https://2011.igem.org/Team:Washington/Protocols/redesign_cell_lysate_assay Cell Lysate Assay by Decarbonylase Redesign Team]
=Break It:  Gluten Destruction Protocols=
=Break It:  Gluten Destruction Protocols=
-
[https://2011.igem.org/Team:Washington/Protocols/Cell_Lysate_Assay Cell Lysate Assay]
+
[https://2011.igem.org/Team:Washington/Protocols/Cell_Lysate_Assay Whole Cell Lysate Assay]
[https://2011.igem.org/Team:Washington/Protocols/50mL_Scale Small Scale (50mL) Protein Expression and Purification]
[https://2011.igem.org/Team:Washington/Protocols/50mL_Scale Small Scale (50mL) Protein Expression and Purification]
 +
[https://2011.igem.org/Team:Washington/Protocols/Purified_Enzyme_Assay Enzyme Assay]
-
=Make It: Magnetosome Protocols=
 
-
[https://2011.igem.org/Team:Washington/Protocols/Gib_Rxn Gibson Reaction]
+
=Make It: iGEM Toolkits=
-
[https://2011.igem.org/Team:Washington/Protocols/Gib_Purif. Gibson Purification]
+
[https://2011.igem.org/Team:Washington/Protocols/Cyto. Cytometry Protocol]
-
=Gibson Vectors (pGB) protocols=
+
[https://2011.igem.org/Team:Washington/Protocols/Elect. Electroporation (Transformation)]
 +
[https://2011.igem.org/Team:Washington/Protocols/Gib_Rxn Gibson Cloning/Assembly]
 +
[https://2011.igem.org/Team:Washington/Protocols/Gib_Purif. Gibson Purification]
 +
[https://2011.igem.org/Team:Washington/Protocols/High_PCR High-Yield PCR (Full-Gene Assembly)]
 +
[https://2011.igem.org/Team:Washington/Protocols/Plas_DNA. Isolation of Plasmid DNA (miniprep)]
 +
[https://2011.igem.org/Team:Washington/Protocols/Induc_studies. Induction Studies of Proteins Fusions (mam-sfGFP)]
 +
[https://2011.igem.org/Team:Washington/Protocols/pGA. pGA Vector Assay]
 +
[https://2011.igem.org/Team:Washington/Protocols/PBS. PBS Stock Protocol]
 +
[https://2011.igem.org/Team:Washington/Protocols/Overnights. Preparation of Overnight Cultures]
-
 
+
=Wiki Design=
-
Check out or add wiki design tools here:
+
[https://2011.igem.org/Team:Washington/Protocols/Wiki_Design Wiki Design Tools (Wiki Markup, WikiDust, etc.)]
-
[https://2011.igem.org/Team:Washington/Protocols/Wiki_Design Wiki Design]
+
-
 
+
-
 
+
-
 
+
-
 
+
-
'''Colony PCR with Green tag'''
+
-
 
+
-
Master mix(7ul):
+
-
 
+
-
1ul 10uM forword primer
+
-
 
+
-
1ul 10uM reverse Primer
+
-
 
+
-
5ul 2x Green tag
+
-
 
+
-
Cell water(3ul): Pick one colony from the plate and mix with 10ul of ddH2O to make 10ul of cell water
+
-
 
+
-
Reaction = Master mix(7ul) + Cell water(3ul) = 10ul per tube
+
-
 
+
-
Use program "Colony" & change the extention time (1min per kb)
+
-
 
+
-
'''Heat Shock Transformation'''
+
-
 
+
-
2 ul ligation
+
-
 
+
-
20 ul cells
+
-
 
+
-
Ice 20 minutes
+
-
 
+
-
Heat shock at 42C for 1 minute
+
-
 
+
-
Ice 2 minutes
+
-
 
+
-
Prepare 200 ul of TB (no anti) and transformed cells in culture tube
+
-
 
+
-
Incubate at 37C for 1 hour
+
-
 
+
-
Plate cells
+

Latest revision as of 02:10, 29 September 2011


Protocols


General Protocols

General Agarose Gel Electrophoresis

General PCR Protocol

General Digestion Protocol

General Ligation Protocol

General Transformation Protocol

Colony PCR Protocol

Competent Cell Prep Protocol

Kunkel Mutagenesis

[http://www.bio.davidson.edu/Courses/molbio/kunkel/kunkel.html Overview of the Kunkel Mutagenesis process]

Standard 1L Expression Purification

Gene Assembly With Oligos

Sequencing

Computational Protein Design

Glycerol Stocks


Make It: Diesel Production Protocols

Alkane Biosynthesis media and extraction

Alkane Biosynthesis cloning

Cell Lysate Assay by Decarbonylase Redesign Team


Break It: Gluten Destruction Protocols

Whole Cell Lysate Assay

Small Scale (50mL) Protein Expression and Purification

Enzyme Assay


Make It: iGEM Toolkits

Cytometry Protocol

Electroporation (Transformation)

Gibson Cloning/Assembly

Gibson Purification

High-Yield PCR (Full-Gene Assembly)

Isolation of Plasmid DNA (miniprep)

Induction Studies of Proteins Fusions (mam-sfGFP)

pGA Vector Assay

PBS Stock Protocol

Preparation of Overnight Cultures


Wiki Design

Wiki Design Tools (Wiki Markup, WikiDust, etc.)

Retrieved from "http://2011.igem.org/Team:Washington/Protocols"