Team:UT-Tokyo/Project/Results
From 2011.igem.org
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{{:Team:UT-Tokyo/Templates/BeginContent|fullpagename=Team:UT-Tokyo/Project|subpagename=Project}} | {{:Team:UT-Tokyo/Templates/BeginContent|fullpagename=Team:UT-Tokyo/Project|subpagename=Project}} | ||
- | == | + | <html> |
+ | <style type="text/css"> | ||
+ | div.float-left{float:left; width:482px;} | ||
+ | div.float-right{float:right; width:482px;} | ||
+ | span.sub{vertical-align: sub;} | ||
+ | span.super{vertical-align:super;} | ||
+ | span.under{text-decoration: underline;} | ||
+ | |||
+ | #top-panel { border-bottom:3px solid #e8f3c6;} | ||
+ | #sub-panel { text-align:center; } | ||
+ | #sub-panel a { width:150px; float:right; color:#000; text-decoration:none; margin-right:30px; font-weight:bold; } | ||
+ | #sub-panel a span { padding:6px; display:block; } | ||
+ | |||
+ | </style> | ||
+ | |||
+ | <script type="text/javascript"> | ||
+ | $(function(){ | ||
+ | $("#sub-panel").click(function(){ | ||
+ | $("#top-panel").slideToggle(); | ||
+ | var el = $("#shText"); | ||
+ | var state = $("#shText").html(); | ||
+ | state = (state == 'Hide' ? '<span id="shText">Show</span>' : '<span id="shText">Hide</span>'); | ||
+ | el.replaceWith(state); | ||
+ | }); | ||
+ | }); | ||
+ | </script> | ||
+ | |||
+ | |||
+ | </html> | ||
+ | |||
+ | ==Assembly parts== | ||
+ | ===Digest=== | ||
+ | |||
+ | ===Ligation=== | ||
+ | |||
+ | ===colony PCR=== | ||
+ | |||
+ | ==Transformation== | ||
+ | ===Making Competent E. coli cell=== | ||
+ | |||
+ | ===Transformation of E. coli=== | ||
+ | <div id="top-panel"> | ||
+ | ====Preparation==== | ||
+ | |||
+ | :iGEM parts / ligation products | ||
+ | :SOC or LB (No antibiotic) 500μL | ||
+ | :TE 15μL | ||
+ | :plates | ||
+ | :ice box | ||
+ | :heat block(42℃) | ||
+ | :competent cells | ||
+ | → always on ice! Melt on ice! Mix DNA as soon as cells melt! | ||
+ | |||
+ | ====Protocol==== | ||
+ | |||
+ | to thaw out igem parts | ||
+ | |||
+ | :#With a pipette tip, punch a hole in the foil | ||
+ | :#Add 15uL of TE (MilliQ),and pipetting | ||
+ | :#Pipette 1uL of the resuspended DNA Transformation into your desired competent cells | ||
+ | :#Hold on ice for 30 mins | ||
+ | :#Heat shock at 42°C for 45 seconds (and on ice after it) | ||
+ | :#Add 300uL of LBborth in each epp | ||
+ | :#Wait for 10 mins | ||
+ | :#Hold at 37℃ for 30 mins | ||
+ | :#Plate out | ||
+ | :#Incubate at 37°C | ||
+ | </div> | ||
{{:Team:UT-Tokyo/Templates/EndContent}} | {{:Team:UT-Tokyo/Templates/EndContent}} |
Revision as of 11:20, 24 September 2011
Project
iGEM UT-Tokyo
Assembly parts
Digest
Ligation
colony PCR
Transformation
Making Competent E. coli cell
Transformation of E. coli
Preparation
- iGEM parts / ligation products
- SOC or LB (No antibiotic) 500μL
- TE 15μL
- plates
- ice box
- heat block(42℃)
- competent cells
→ always on ice! Melt on ice! Mix DNA as soon as cells melt!
Protocol
to thaw out igem parts
- With a pipette tip, punch a hole in the foil
- Add 15uL of TE (MilliQ),and pipetting
- Pipette 1uL of the resuspended DNA Transformation into your desired competent cells
- Hold on ice for 30 mins
- Heat shock at 42°C for 45 seconds (and on ice after it)
- Add 300uL of LBborth in each epp
- Wait for 10 mins
- Hold at 37℃ for 30 mins
- Plate out
- Incubate at 37°C