From 2011.igem.org

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 {{Template:Team:Washington/Templates/Top}}
+__NOTOC__
-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]
-[http://soslab.ee.washington.edu/igem/2011/index.php/Sequencing  Sequencing Instructions]
-[http://soslab.ee.washington.edu/igem/2011/index.php/KunkelMutagensis Kunkel Mutagensis]
+[https://2011.igem.org/Team:Washington/Protocols/PCR  General PCR Protocol]
-[http://www.bio.davidson.edu/Courses/molbio/kunkel/kunkel.html Overview of how Kunkel Mutagensis works]
+[https://2011.igem.org/Team:Washington/Protocols/Digestion  General Digestion Protocol]
-[http://soslab.ee.washington.edu/igem/2011/index.php/PlateExpression  96 Well Plate Protein Expression]
+[https://2011.igem.org/Team:Washington/Protocols/Ligation  General Ligation Protocol]
-[http://soslab.ee.washington.edu/igem/2011/index.php/DoubleDigest Double Digest]
+[https://2011.igem.org/Team:Washington/Protocols/Transformation  General Transformation Protocol]
-[http://soslab.ee.washington.edu/igem/2011/index.php/Projects:Biofuels/AlkaneBiosynthesis Alkane Biosynthesis Protocol]
+[https://2011.igem.org/Team:Washington/Protocols/Colony Colony PCR 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/Competent  Competent Cell Prep Protocol]
-[http://soslab.ee.washington.edu/igem/2011/index.php/Protocols/GeneAssembly Gene Assembly With Oligos]
+[https://2011.igem.org/Team:Washington/Protocols/Kunkel Kunkel Mutagenesis]
+[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]
-=Protocol Page=
+[https://2011.igem.org/Team:Washington/Protocols/gene_assembly Gene Assembly With Oligos]
-'''restriction digest'''
+[https://2011.igem.org/Team:Washington/Protocols/sequencing Sequencing]
-uL DNA
-uL buffer ( 2 for most, check NEB)
+[https://2011.igem.org/Team:Washington/Protocols/CompDesign  Computational Protein Design]
-.5 uL BSA
+[https://2011.igem.org/Team:Washington/Protocols/Glycerol_Stocks  Glycerol Stocks]
-uL enzyme 1
-uL enzyme 2
+=Make It:  Diesel Production Protocols=
-water to 50 uL(32.5 uL, add first)
+[https://2011.igem.org/Team:Washington/alkanebiosynthesis Alkane Biosynthesis media and extraction]
-'''oligo assembly by PCR'''
+[https://2011.igem.org/Team:Washington/alkanebiosynthesis_cloning Alkane Biosynthesis cloning]
-resuspend oligos with water, amount of water= concentration(in nm)*10 in uL
+[https://2011.igem.org/Team:Washington/Protocols/redesign_cell_lysate_assay Cell Lysate Assay by Decarbonylase Redesign Team]
-make oligo mix with 5uL of each primer
-PCR reaction:
+=Break It:  Gluten Destruction Protocols=
-uL phusion
-.5uL oligo mix
+[https://2011.igem.org/Team:Washington/Protocols/Cell_Lysate_Assay Whole Cell Lysate Assay]
-uL first oligo
+[https://2011.igem.org/Team:Washington/Protocols/50mL_Scale Small Scale (50mL) Protein Expression and Purification]
-uL last oligo
+[https://2011.igem.org/Team:Washington/Protocols/Purified_Enzyme_Assay Enzyme Assay]
-uL buffer
-uL dnTP
+=Make It: iGEM Toolkits=
-dH20 to 50uL
+[https://2011.igem.org/Team:Washington/Protocols/Cyto. Cytometry Protocol]
-'''Ligation'''
+[https://2011.igem.org/Team:Washington/Protocols/Elect. Electroporation (Transformation)]
-uL insert
+[https://2011.igem.org/Team:Washington/Protocols/Gib_Rxn Gibson Cloning/Assembly]
-uL vector
+[https://2011.igem.org/Team:Washington/Protocols/Gib_Purif. Gibson Purification]
-uL T4 ligase buffer
+[https://2011.igem.org/Team:Washington/Protocols/High_PCR High-Yield PCR (Full-Gene Assembly)]
-uL T4 ligase
+[https://2011.igem.org/Team:Washington/Protocols/Plas_DNA. Isolation of Plasmid DNA (miniprep)]
-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/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]
-'''Colony PCR with Green tag'''
+[https://2011.igem.org/Team:Washington/Protocols/Overnights. Preparation of Overnight Cultures]
-Master mix(7ul):
-ul 10uM forword primer
-ul 10uM reverse Primer
+=Wiki Design=
+[https://2011.igem.org/Team:Washington/Protocols/Wiki_Design Wiki Design Tools (Wiki Markup, WikiDust, etc.)]
-ul 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'''
-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