IGEM:IMPERIAL/2008/New/Protocols: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
mNo edit summary
mNo edit summary
 
(28 intermediate revisions by 3 users not shown)
Line 1: Line 1:
{{Imperial/StartPage2}}
{{Imperial/StartPage2}}
= Protocols =


{| cellpadding="1" style="background:#66aadd; border:1px solid #66aadd; color:black" align="left" width=75%
 
|width="100%"| <font size=4px><font color=white>'''1. Transformation Protocols for integration vectors in ''Bacillus subtilis'''''
{{Imperial/Box2|Experiments & Protocols|
Below you will find a comprehensive list of all the protocols we used during our wet lab work. We spent some time toward the start of the project sourcing and updating effective protocols for ''B. subtilis'' manipulation explicitly, as other teams have previously had difficulty in this area.
 
These protocols worked for us, so any future teams looking to use ''subtilis'' could save quite a bit of time by looking here before diving into the literature!}}
 
 
{| cellpadding="1" style="background:#66aadd; border:1px solid #66aadd; color:black" align="center" width=50%
|width="100%"| <font size=4px color=white>'''1. Transformation Protocols for integration vectors in ''Bacillus subtilis'''''</font>
|-
|-
|style="background:#FFFFFF"|  
|style="background:#FFFFFF"|  
Line 9: Line 15:
*[[/Transformation_protocol_4 | Protocol 2]]
*[[/Transformation_protocol_4 | Protocol 2]]
|-  
|-  
|style="background:#66aadd"| <font size=4px><font color=white>'''2. Transformation Protocol for linear DNA in ''Bacillus subtilis'''''
|style="background:#66aadd"| <font size=4px color=white>'''2. Transformation Protocol for linear DNA in ''Bacillus subtilis'''''</font>
|-
|-
|style="background:#FFFFFF"|  
|style="background:#FFFFFF"|  
*Protocol
*Protocol
|-  
|-  
|style="background:#66aadd"| <font size=4px><font color=white>'''3. Transformation Protocol ''E.coli'''''
|style="background:#66aadd"| <font size=4px color=white>'''3. Transformation Protocol ''E.coli'''''</font>
|-
|-
|style="background:#FFFFFF"|  
|style="background:#FFFFFF"|  
*[[/XL1-Blue_preparation | Preparation of XL1-blue electrocompetent cells]]
*[[/XL1-Blue_preparation | Preparation of XL1-blue electrocompetent cells]]
|-  
|-  
|style="background:#66aadd"| <font size=4px><font color=white>'''4. Motility Analysis'''
|style="background:#66aadd"| <font size=4px color=white>'''4. Motility Analysis'''</font>
|-
|-
|style="background:#FFFFFF"|  
|style="background:#FFFFFF"|  
*[[/Protocols#Motility_Experiments | Motility Assays]]
*[[/Protocols#Motility_Experiments | Motility Assays]]
|-  
|-  
|style="background:#66aadd"| <font size=4px><font color=white>'''5. PCR'''
|style="background:#66aadd"| <font size=4px color=white>'''5. PCR'''</font>
|-
|-
|style="background:#FFFFFF"|  
|style="background:#FFFFFF"|  
Line 30: Line 36:
*[[/Colony_PCR| Single Colony PCR]]
*[[/Colony_PCR| Single Colony PCR]]
|-  
|-  
|style="background:#66aadd"| <font size=4px><font color=white>'''6. Calibration Curves'''
|style="background:#66aadd"| <font size=4px color=white>'''6. Calibration Curves'''</font>
|-
|-
|style="background:#FFFFFF"|  
|style="background:#FFFFFF"|  
Line 37: Line 43:
*[[/Fluorescence| Fluorescence vs GFPmut3b]]
*[[/Fluorescence| Fluorescence vs GFPmut3b]]
|-  
|-  
|style="background:#66aadd"| <font size=4px><font color=white>'''7. Promoter-RBS Testing Protocols'''
|style="background:#66aadd"| <font size=4px color=white>'''7. Promoter-RBS Testing Protocols'''</font>
|-
|-
|style="background:#FFFFFF"|  
|style="background:#FFFFFF"|  
Line 44: Line 50:
*[[/Light_Inducible | Light Inducible Promoters and RBS]]
*[[/Light_Inducible | Light Inducible Promoters and RBS]]
|-  
|-  
|style="background:#66aadd"| <font size=4px><font color=white>'''8. Biomaterial Protocols'''
|style="background:#66aadd"| <font size=4px color=white>'''8. Biomaterial Protocols'''</font>
|-
|-
|style="background:#FFFFFF"|  
|style="background:#FFFFFF"|  
*Protocol
*Protocol
|}
|}
 
<br clear="all"><br><hr>
{{Imperial/EndPage}}
{{Imperial/EndPage|Cloning_Strategy|Major_Results}}

Latest revision as of 05:20, 30 September 2008

<html> <style type="text/css"> .firstHeading {display: none;} </style> </html> <html> <style type="text/css"> 

   table.calendar          { margin:0; padding:2px; }

table.calendar td { margin:0; padding:1px; vertical-align:top; } table.month .heading td { padding:1px; background-color:#FFFFFF; text-align:center; font-size:120%; font-weight:bold; } table.month .dow td { text-align:center; font-size:110%; } table.month td.today { background-color:#3366FF } table.month td { 

   border:2px;
   margin:0;
   padding:0pt 1.5pt;
   font-size:8pt;
   text-align:right;
   background-color:#FFFFFF;
   }
  1. bodyContent table.month a { background:none; padding:0 }

.day-active { font-weight:bold; } .day-empty { color:black; } </style> </html>

<html><script language="JavaScript">

var timeout = 250; var closetimer = 0; var ddmenuitem = 0;

// open hidden layer function mopen(id) { // cancel close timer mcancelclosetime(); // close old layer if(ddmenuitem) ddmenuitem.style.visibility = 'hidden'; // get new layer and show it ddmenuitem = document.getElementById(id); ddmenuitem.style.visibility = 'visible'; } // close showed layer function mclose() { if(ddmenuitem) ddmenuitem.style.visibility = 'hidden'; } // go close timer function mclosetime() { closetimer = window.setTimeout(mclose, timeout); } // cancel close timer function mcancelclosetime() { if(closetimer) { window.clearTimeout(closetimer); closetimer = null; } } // close layer when click-out //document.onclick = mclose; </script> <table background="http://i59.photobucket.com/albums/g305/Timpski/ToolbarBackground.png" style="color:#ffffff;" link="#ffffff" cellpadding="0" cellspacing="1" border="0" bordercolor="#ffffff" align="center" width="100%"><tr><td colspan="6" class="wetlab"><br><br><br></td></tr> <tr><td align="center" width="10%" valign="bottom"><ul id="sddm"><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Home"> Home </a></ul> </td><td align="center" width="20%" valign="bottom"><ul id="sddm"><a href="#" 

       onclick="mopen('m1')" 
       onmouseover="mopen('m1')" 
       onmouseout="mclosetime()">Biofabricator Subtilis</a>
       <div id="m1" 
           onmouseover="mcancelclosetime()" 
           onmouseout="mclosetime()">
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Project">Project Specifications</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Chassis_1">Why B. subtilis?</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Chassis_2">B. subtilis: Benefits vs Challenges</a>
       </div></ul>

</td><td align="center" width="18%" valign="bottom"><ul id="sddm"><a href="#" 

       onclick="mopen('m2')" 
       onmouseover="mopen('m2')" 
       onmouseout="mclosetime()">Wet Lab</a>
       <div id="m2" 
           onmouseover="mcancelclosetime()" 
           onmouseout="mclosetime()">
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Wet_Lab">Wet Lab Hub</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Cloning_Strategy">Cloning Strategy</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Protocols">Experiments & Protocols</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Major_Results">Experimental Results</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/BioBricks">BioBricks & Characterisation</a>
       </div></ul>

</td><td align="center" width="18%" valign="bottom"><ul id="sddm"><a href="#" 

       onclick="mopen('m3')" 
       onmouseover="mopen('m3')" 
       onmouseout="mclosetime()">Dry Lab</a>
       <div id="m3" 
           onmouseover="mcancelclosetime()" 
           onmouseout="mclosetime()">
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Dry_Lab">Dry Lab Hub</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Growth_Curve">Growth Curves</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Genetic_Circuit">Genetic Circuits</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Motility">Motility Analysis</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Appendices">Appendices - Code etc.</a>
       </div></ul>

</td><td align="center" width="17%" valign="bottom"><ul id="sddm"><a href="http://2008.igem.org/Team:Imperial_College/Notebook"> Notebook </a></ul> </td><td align="center" width="17%" valign="bottom"><ul id="sddm"><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Team"> Our Team </a></ul> </td></tr></table></html>

<html><style type="text/css">

div.Section { font:11pt/16pt Calibri, Verdana, Arial, Geneva, sans-serif; background-image: url(http://openwetware.org/images/a/a0/Background.PNG); background-size: 100%; background-origin: content; }

/* Text (paragraphs) */ div.Section p { font:11pt/16pt Calibri, Verdana, Arial, Geneva, sans-serif; text-align:justify; margin-top:0px; margin-left:30px; margin-right:30px; }

/* Headings */ div.Section h1 { font:22pt Calibri, Verdana, Arial, Geneva, sans-serif; text-align:left; color:#3366FF; font-weight:bold; }

/* Subheadings */ div.Section h2 { font:18pt Calibri, Verdana, Arial, Geneva, sans-serif; color:#3366FF; margin-left:5px; font-weight:bold; }

/* Subsubheadings */ div.Section h3 { font:22pt Calibri, Verdana, Arial, sans-serif; color:#E5EBFF; margin-left:10px; font-weight:bold; }

/* Subsubsubheadings */ div.Section h4 { font:22pt Calibri, Verdana, Arial, sans-serif; color:#2B48B3; margin-left:10px; font-weight:bold; }

/* Subsubsubsubheadings */ div.Section h5 { font:12pt Calibri, Verdana, Arial, sans-serif; color:#3366FF; margin-left:20px; }

/* References */ div.Section h6 { font:12pt Calibri, Verdana, Arial, sans-serif; font-weight:bold; font-style:italic; color:#3366FF; margin-left:25px; }

/* Hyperlinks */ div.Section a { }

div.Section a:hover { }

/* Tables */ div.Section td { font:11pt/16pt Calibri, Verdana, Arial, Geneva, sans-serif; text-align:justify; vertical-align:top; padding:2px 4px 2px 4px; }

/* Lists */ div.Section li { font:11pt/16pt Calibri, Verdana, Arial, Geneva, sans-serif; text-align:left; margin-top:0px; margin-left:30px; margin-right:0px; }

/* TOC stuff */ table.toc { margin-left:10px; }

table.toc li { font: 11pt/16pt Calibri, Verdana, Arial, Geneva, sans-serif; text-align: justify; margin-top: 0px; margin-left:2px; margin-right:2px; }

/* [edit] links */ span.editsection { color:#BBBBBB; font-size:10pt; font-weight:normal; font-style:normal; vertical-align:bottom; }

span.editsection a { color:#BBBBBB; font-size:10pt; font-weight:normal; font-style:normal; vertical-align:bottom; }

span.editsection a:hover { color:#3366FF; font-size:10pt; font-weight:normal; font-style:normal; vertical-align:bottom; }

/* Drop-down Menu */

  1. sddm {

margin: 0; padding: 0; z-index: 30 margin: 0; padding: 0; float: center; font: bold 12pt Calibri, Verdana, Arial, Geneva, sans-serif; border: 0px; list-style: none; }

  1. sddm a {

display: block; margin: 0px 0px 0px 0px; padding: 0 0 12px 0; color: #FFFFFF; text-align: center; text-decoration: none; }

  1. sddm a:hover {

border: 0px }

  1. sddm div {

position: absolute; visibility: hidden; margin: 0; padding: 0; background: #66aadd; border: 1px solid #66aadd } #sddm div a { position: relative; left: 0; display: block; margin: 0; padding: 5px 10px; width: auto; white-space: nowrap; text-align: left; text-decoration: none; background: #FFFFFF; color: #2875DE; font: 11pt Calibri, Verdana, Arial, Geneva, sans-serif } #sddm div a:hover { background: #66aadd; color: #FFFFFF } </style></html>


Experiments & Protocols

Below you will find a comprehensive list of all the protocols we used during our wet lab work. We spent some time toward the start of the project sourcing and updating effective protocols for B. subtilis manipulation explicitly, as other teams have previously had difficulty in this area.

These protocols worked for us, so any future teams looking to use subtilis could save quite a bit of time by looking here before diving into the literature!



1. Transformation Protocols for integration vectors in Bacillus subtilis
2. Transformation Protocol for linear DNA in Bacillus subtilis
  • Protocol
3. Transformation Protocol E.coli
4. Motility Analysis
5. PCR
6. Calibration Curves
7. Promoter-RBS Testing Protocols
8. Biomaterial Protocols
  • Protocol



<html><center><table style="color:#ffffff;background-color:#66aadd;" cellpadding="3" cellspacing="1" border="0" bordercolor="#ffffff" align="center"> <tr><td><ul id="sddm"></html>< Previous<html></ul> </td><td><ul id="sddm"><a href="#">Back to top</a></ul> </td><td><ul id="sddm"></html>Next ><html></ul> </td></tr></table> </center></html>

Retrieved from "https://openwetware.org/mediawiki/index.php?title=IGEM:IMPERIAL/2008/New/Protocols&oldid=247578"