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