From 2011.igem.org

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 {{Template:Team:Washington/Templates/Top}}
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-INSERT INFO HERE... LOOK AT OTHER PAGES FOR EXAMPLES OF FORMATING AND INSERTING PICTURES.   CLICK THE EDIT BUTTON ON THE UPPER LEFT SIDE OF THE PAGE AFTER YOU HAVE SIGNED IN.
+<center><big><big><big><big>'''Protocols'''</big></big></big></big></center><br><br>
-[https://2011.igem.org/Team:Washington/Alkanes/Protocols/Example_1 Example 1]
+=General Protocols=
-[https://2011.igem.org/Team:Washington/Alkanes/Protocols/Example_2 Example 2]
+[https://2011.igem.org/Team:Washington/Protocols/gel_electrophoresis  General Agarose Gel Electrophoresis]
+[https://2011.igem.org/Team:Washington/Protocols/PCR  General PCR Protocol]
-[http://soslab.ee.washington.edu/igem/2011/index.php/KunkelMutagensis Kunkel Mutagensis]
+[https://2011.igem.org/Team:Washington/Protocols/Digestion  General Digestion Protocol]
-[http://www.bio.davidson.edu/Courses/molbio/kunkel/kunkel.html Overview of how Kunkel Mutagensis works]
+[https://2011.igem.org/Team:Washington/Protocols/Ligation  General Ligation Protocol]
-[http://soslab.ee.washington.edu/igem/2011/index.php/PlateExpression  96 Well Plate Protein Expression]
+[https://2011.igem.org/Team:Washington/Protocols/Transformation  General Transformation Protocol]
-[http://soslab.ee.washington.edu/igem/2011/index.php/DoubleDigest Double Digest]
+[https://2011.igem.org/Team:Washington/Protocols/Colony Colony PCR Protocol]
-[http://soslab.ee.washington.edu/igem/2011/index.php/Projects:Biofuels/AlkaneBiosynthesis Alkane Biosynthesis Protocol]
+[https://2011.igem.org/Team:Washington/Protocols/Competent  Competent Cell Prep Protocol]
-[http://soslab.ee.washington.edu/igem/2011/index.php/Protocols/1L_NiNTA_Expression_Purification Standard 1L Expression Purification]
+[https://2011.igem.org/Team:Washington/Protocols/Kunkel Kunkel Mutagenesis]
-[http://soslab.ee.washington.edu/igem/2011/index.php/Protocols/GeneAssembly Gene Assembly With Oligos]
+[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/gene_assembly Gene Assembly With Oligos]
-=Protocol Page=
+[https://2011.igem.org/Team:Washington/Protocols/sequencing Sequencing]
-'''restriction digest'''
+[https://2011.igem.org/Team:Washington/Protocols/CompDesign  Computational Protein Design]
-uL DNA
-uL buffer ( 2 for most, check NEB)
+[https://2011.igem.org/Team:Washington/Protocols/Glycerol_Stocks  Glycerol Stocks]
-.5 uL BSA
-uL enzyme 1
+=Make It:  Diesel Production Protocols=
-uL enzyme 2
+[https://2011.igem.org/Team:Washington/alkanebiosynthesis Alkane Biosynthesis media and extraction]
-water to 50 uL(32.5 uL, add first)
+[https://2011.igem.org/Team:Washington/alkanebiosynthesis_cloning Alkane Biosynthesis cloning]
-'''oligo assembly by PCR'''
+[https://2011.igem.org/Team:Washington/Protocols/redesign_cell_lysate_assay Cell Lysate Assay by Decarbonylase Redesign Team]
-resuspend oligos with water, amount of water= concentration(in nm)*10 in uL
-make oligo mix with 5uL of each primer
+=Break It:  Gluten Destruction Protocols=
-PCR reaction:
+[https://2011.igem.org/Team:Washington/Protocols/Cell_Lysate_Assay Whole Cell Lysate Assay]
-uL phusion
-.5uL oligo mix
+[https://2011.igem.org/Team:Washington/Protocols/50mL_Scale Small Scale (50mL) Protein Expression and Purification]
-uL first oligo
+[https://2011.igem.org/Team:Washington/Protocols/Purified_Enzyme_Assay Enzyme Assay]
-uL last oligo
-uL buffer
+=Make It: iGEM Toolkits=
-uL dnTP
+[https://2011.igem.org/Team:Washington/Protocols/Cyto. Cytometry Protocol]
-dH20 to 50uL
+[https://2011.igem.org/Team:Washington/Protocols/Elect. Electroporation (Transformation)]
-'''Ligation'''
+[https://2011.igem.org/Team:Washington/Protocols/Gib_Rxn Gibson Cloning/Assembly]
-uL insert
+[https://2011.igem.org/Team:Washington/Protocols/Gib_Purif. Gibson Purification]
-uL vector
+[https://2011.igem.org/Team:Washington/Protocols/High_PCR High-Yield PCR (Full-Gene Assembly)]
-uL T4 ligase buffer
+[https://2011.igem.org/Team:Washington/Protocols/Plas_DNA. Isolation of Plasmid DNA (miniprep)]
-uL T4 ligase
+[https://2011.igem.org/Team:Washington/Protocols/Induc_studies. Induction Studies of Proteins Fusions (mam-sfGFP)]
-incubate at <s>37C</s> no.  **Note that NEB website recommends 16C - room temp for ligations, we do ours at RT and it works well.  1 hour.
+[https://2011.igem.org/Team:Washington/Protocols/pGA. pGA Vector Assay]
-'''Colony PCR with Green tag'''
+[https://2011.igem.org/Team:Washington/Protocols/PBS. PBS Stock Protocol]
-Master mix(7ul):
+[https://2011.igem.org/Team:Washington/Protocols/Overnights. Preparation of Overnight Cultures]
-ul 10uM forword primer
-ul 10uM reverse Primer
-ul 2x Green tag
+=Wiki Design=
+[https://2011.igem.org/Team:Washington/Protocols/Wiki_Design Wiki Design Tools (Wiki Markup, WikiDust, etc.)]
-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'''
-ul ligation
-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