From 2011.igem.org

@@ Line 1: / Line 1: @@
 {{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]
-=Protocol Page=
-'''restriction digest'''
+[https://2011.igem.org/Team:Washington/Protocols/PCR  General PCR Protocol]
-uL DNA
-uL buffer ( 2 for most, check NEB)
+[https://2011.igem.org/Team:Washington/Protocols/Digestion  General Digestion Protocol]
-.5 uL BSA
+[https://2011.igem.org/Team:Washington/Protocols/Ligation  General Ligation Protocol]
-uL enzyme 1
+[https://2011.igem.org/Team:Washington/Protocols/Transformation  General Transformation Protocol]
-uL enzyme 2
+[https://2011.igem.org/Team:Washington/Protocols/Colony Colony PCR Protocol]
-water to 50 uL(32.5 uL, add first)
+[https://2011.igem.org/Team:Washington/Protocols/Competent  Competent Cell Prep Protocol]
-'''oligo assembly by PCR'''
+[https://2011.igem.org/Team:Washington/Protocols/Kunkel Kunkel Mutagenesis]
-resuspend oligos with water, amount of water= concentration(in nm)*10 in uL
+[http://www.bio.davidson.edu/Courses/molbio/kunkel/kunkel.html Overview of the Kunkel Mutagenesis process]
-make oligo mix with 5uL of each primer
+[https://2011.igem.org/Team:Washington/Protocols/expression_purification Standard 1L Expression Purification]
-PCR reaction:
+[https://2011.igem.org/Team:Washington/Protocols/gene_assembly Gene Assembly With Oligos]
-uL phusion
-.5uL oligo mix
+[https://2011.igem.org/Team:Washington/Protocols/sequencing Sequencing]
-uL first oligo
+[https://2011.igem.org/Team:Washington/Protocols/CompDesign  Computational Protein Design]
-uL last oligo
+[https://2011.igem.org/Team:Washington/Protocols/Glycerol_Stocks  Glycerol Stocks]
-uL buffer
-uL dnTP
+=Make It:  Diesel Production Protocols=
-dH20 to 50uL
+[https://2011.igem.org/Team:Washington/alkanebiosynthesis Alkane Biosynthesis media and extraction]
-'''Ligation'''
+[https://2011.igem.org/Team:Washington/alkanebiosynthesis_cloning Alkane Biosynthesis cloning]
-uL insert
+[https://2011.igem.org/Team:Washington/Protocols/redesign_cell_lysate_assay Cell Lysate Assay by Decarbonylase Redesign Team]
-uL vector
-uL T4 ligase buffer
+=Break It:  Gluten Destruction Protocols=
-uL T4 ligase
+[https://2011.igem.org/Team:Washington/Protocols/Cell_Lysate_Assay Whole Cell Lysate 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/50mL_Scale Small Scale (50mL) Protein Expression and Purification]
+[https://2011.igem.org/Team:Washington/Protocols/Purified_Enzyme_Assay Enzyme Assay]
-'''Colony PCR with Green tag'''
+=Make It: iGEM Toolkits=
-Master mix(7ul):
+[https://2011.igem.org/Team:Washington/Protocols/Cyto. Cytometry Protocol]
-ul 10uM forword primer
+[https://2011.igem.org/Team:Washington/Protocols/Elect. Electroporation (Transformation)]
-ul 10uM reverse Primer
+[https://2011.igem.org/Team:Washington/Protocols/Gib_Rxn Gibson Cloning/Assembly]
-ul 2x Green tag
+[https://2011.igem.org/Team:Washington/Protocols/Gib_Purif. Gibson Purification]
-Cell water(3ul): Pick one colony from the plate and mix with 10ul of ddH2O to make 10ul of cell water
+[https://2011.igem.org/Team:Washington/Protocols/High_PCR High-Yield PCR (Full-Gene Assembly)]
-Reaction = Master mix(7ul) + Cell water(3ul) = 10ul per tube
+[https://2011.igem.org/Team:Washington/Protocols/Plas_DNA. Isolation of Plasmid DNA (miniprep)]
-Use program "Colony" & change the extention time (1min per kb)
+[https://2011.igem.org/Team:Washington/Protocols/Induc_studies. Induction Studies of Proteins Fusions (mam-sfGFP)]
-'''Heat Shock Transformation'''
+[https://2011.igem.org/Team:Washington/Protocols/pGA. pGA Vector Assay]
-ul ligation
+[https://2011.igem.org/Team:Washington/Protocols/PBS. PBS Stock Protocol]
-ul cells
+[https://2011.igem.org/Team:Washington/Protocols/Overnights. Preparation of Overnight Cultures]
-Ice 20 minutes
-Heat shock at 42C for 1 minute
-Ice 2 minutes
+=Wiki Design=
+[https://2011.igem.org/Team:Washington/Protocols/Wiki_Design Wiki Design Tools (Wiki Markup, WikiDust, etc.)]
-Prepare 200 ul of TB (no anti) and transformed cells in culture tube
-Incubate at 37C for 1 hour
-Plate cells