Team:TU Darmstadt/protocols/Cell counting/plating

From 2013.igem.org

(Difference between revisions)
 
(5 intermediate revisions not shown)
Line 2: Line 2:
<style type="text/css">
<style type="text/css">
-
body
+
body  
{
{
margin:0;  
margin:0;  
Line 15: Line 15:
{
{
   background-color: white;
   background-color: white;
-
   background-image: url("/wiki/images/7/70/Hintergrund3.jpg");
+
   background-image: url("/wiki/images/5/5c/Hintergrund_Grün(ohne_Header%2C_Logo_und_Schlagschatten).jpg");
   background-attachment: scroll;
   background-attachment: scroll;
   background-position: 50% 0;
   background-position: 50% 0;
Line 26: Line 26:
{
{
     background: white;
     background: white;
-
     background-image: url('/wiki/images/2/2f/Header_(mit_Logo).jpg');
+
     background-image: url("/wiki/images/2/2f/Header_%28mit_Logo%29.jpg");
     margin: 0 auto;
     margin: 0 auto;
     height:250px ;
     height:250px ;
Line 35: Line 35:
     background-repeat: no-repeat;
     background-repeat: no-repeat;
     background-size: 100% 100% ;
     background-size: 100% 100% ;
-
     border-style: none;
+
     border-style: none;;}
-
}
+
#p-logo { display:none;}
#p-logo { display:none;}
Line 42: Line 41:
p {line-height:1.5em; margin:0 0 15px; text-align:justify; color:white;}
p {line-height:1.5em; margin:0 0 15px; text-align:justify; color:white;}
h1 {font-size:1.8em; font-weight:400; margin:0 0 12px;}
h1 {font-size:1.8em; font-weight:400; margin:0 0 12px;}
 +
#mm_icon1
#mm_icon1
{
{
position: absolute;
position: absolute;
-
top: 240px;  
+
top: 150px;  
-
left: 200px;
+
left: 30px;
}
}
#mm_icon2
#mm_icon2
{
{
-
position: relative;
+
position: absolute;
-
top: 170px;  
+
top: 150px;  
left: 350px;
left: 350px;
}
}
Line 67: Line 67:
{
{
position: absolute;
position: absolute;
-
top: 260px;  
+
top: 150px;  
-
left: 230px;
+
left: 30px;
background:white;
background:white;
filter:alpha(opacity=83); opacity:0.83;
filter:alpha(opacity=83); opacity:0.83;
Line 80: Line 80:
{
{
position: absolute;
position: absolute;
-
top: 670px;  
+
top: 150px;  
left: 350px;
left: 350px;
background:white;
background:white;
Line 89: Line 89:
border-radius:15px;
border-radius:15px;
}
}
 +
 +
#abstracticon3
#abstracticon3
Line 94: Line 96:
position: absolute;
position: absolute;
top: 150px;  
top: 150px;  
-
left: 700px;
+
left: 670px;
background:white;
background:white;
filter:alpha(opacity=83); opacity:0.83;
filter:alpha(opacity=83); opacity:0.83;
Line 102: Line 104:
border-radius:15px;
border-radius:15px;
}
}
-
 
#taskbar
#taskbar
Line 108: Line 109:
position:absolute;
position:absolute;
top:10px;
top:10px;
-
left:400px;
+
left:500px;
z-index: 5;
z-index: 5;
 +
}
 +
 +
 +
 +
 +
dl.igemTUD2013gelpicture
 +
{
 +
border: 1px solid #000;
 +
background-color: #109f71;
 +
width: 110px;
 +
text-align: center;
 +
padding: 5px 5px 5px 5px;
 +
float: right;
 +
margin: 0 0 0 0;
 +
margin-left:15px
 +
}
 +
 +
.igemTUD2013gelpicture dt
 +
{
 +
font-weight: bold;
 +
background-color: #131210;
 +
color: #959289;
 +
padding: 0 0;
 +
margin-bottom: 10px;
 +
}
 +
 +
.igemTUD2013gelpicture dd img
 +
{
 +
border: 1px solid #000;
 +
width: 100px;
 +
height: 200px;
 +
}
 +
 +
.igemTUD2013gelpicture dd
 +
{
 +
margin: 0;
 +
padding: 5px 5px 5px 5px;
 +
font-size: 100%;
 +
text-align: left;
 +
}
 +
 +
 +
 +
 +
dl.igemTUD2013gelpicture2
 +
{
 +
border: 1px solid #000;
 +
background-color: #109f71;
 +
width: 210px;
 +
text-align: center;
 +
padding: 5px 5px 5px 5px;
 +
float: right;
 +
margin: 0 0 0 0;
 +
margin-left:15px
 +
}
 +
 +
.igemTUD2013gelpicture2 dt
 +
{
 +
font-weight: bold;
 +
background-color: #131210;
 +
color: #959289;
 +
padding: 0 0;
 +
margin-bottom: 10px;
 +
}
 +
 +
.igemTUD2013gelpicture2 dd img
 +
{
 +
border: 1px solid #000;
 +
width: 200px;
 +
height: 200px;
 +
}
 +
 +
.igemTUD2013gelpicture2 dd
 +
{
 +
margin: 0;
 +
padding: 5px 5px 5px 5px;
 +
font-size: 100%;
 +
}
 +
LI.list1 {list-style: none; color:white;}
 +
.blacktext {color:black}
 +
p, ul {
 +
    padding: 0;
 +
    margin: 0;
}
}
</style>
</style>
 +
 +
<body>
 +
 +
<center>
 +
<!-- central main menu -->
 +
 +
<br>
 +
<br>
<!-- Taskbar -->
<!-- Taskbar -->
Line 122: Line 214:
<img alt="Problem" src="/wiki/images/6/66/Darmstadt_green_Problem.jpg" width="100" height="30"></a>
<img alt="Problem" src="/wiki/images/6/66/Darmstadt_green_Problem.jpg" width="100" height="30"></a>
-
<a href="https://2013.igem.org/Team:TU_Darmstadt/solution">
 
-
<img alt="solution" src="/wiki/images/8/87/Darmstadt_green_Solution.jpg" width="100" height="30"></a>
 
<a href="https://2013.igem.org/Team:TU_Darmstadt/result">
<a href="https://2013.igem.org/Team:TU_Darmstadt/result">
Line 145: Line 235:
<a href="https://2013.igem.org/Team:TU_Darmstadt/labbook">
<a href="https://2013.igem.org/Team:TU_Darmstadt/labbook">
-
<img alt="team" src="/wiki/images/f/f3/Darmstadt_green_Labbook.jpg" width="90" height="30"></a>
+
<img alt="team" src="/wiki/images/4/4c/10._Labbook_(angewählt).jpg" width="90" height="30"></a>
</div>
</div>
-
<!-- Cell counting/plating -->
+
 
 +
<br><br><br><br>
<center>
<center>
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/labbook">
 +
<font size="8" color="#F0F8FF" face="Arial regular">Lab book |</font>
 +
</a>
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/materials">
 +
<font size="8" color="#F0F8FF" face="Arial regular">Materials |</font>
 +
</a>
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/protocols">
 +
<font size="8" color="#F0F8FF" face="Arial regular">Protocols</font>
 +
</a>
 +
</center>
<br>
<br>
-
<br>
+
 
-
<br>
+
<!-- Cell counting/plating -->
-
<br>
+
<center>
 +
 
 +
 
<h2><font size="6" color="#F0F8FF" face="Arial regular"> Cell counting/plating </font></h2>
<h2><font size="6" color="#F0F8FF" face="Arial regular"> Cell counting/plating </font></h2>
<div id="all">
<div id="all">
<body>
<body>
-
<font size="3" color="#F0F8FF" face="Arial regular">
+
<p text-aligne:left style="margin-left:50px; margin-right:50px"><font size="4.5" color="#F0F8FF" face="Arial regular">
-
<p text-aligne:left style="margin-left:50px; margin-right:50px">
+
<B> Materials<br></B></font></p>
-
<B> Materials<br></B>
+
<p text-aligne:left style="margin-left:60px; margin-right:50px"><font size="3" color="#F0F8FF" face="Arial regular">
-
LB agar plate<br>
+
 
-
LB media<br>
+
<div align="left" style="margin-left:60px; margin-right:50px">
-
Tubes<br>
+
<ul>
 +
<li class=list1>- LB agar plate</li>
 +
<li class=list1>- <a href="https://2013.igem.org/Team:TU_Darmstadt/materials/Media"> LB media</a></li>
 +
<li class=list1>- Tubes</li>
 +
</ul>
 +
</div>
 +
</font>
 +
</p>
<br>
<br>
-
<B> Procedure<br></B>
+
<p text-aligne:left style="margin-left:50px; margin-right:50px"><font size="4.5" color="#F0F8FF" face="Arial regular">
-
1. Fill each tube in the dilution with 90 &mu;l of LB<br>
+
<B> Procedure<br></B></font></p>
-
2. Add 10 &mu;l of the sample to the first tube and mix<br>
+
<p text-aligne:left style="margin-left:60px; margin-right:60px"><font size="3" color="#F0F8FF" face="Arial regular">
-
3. From the first tube, remove 10 &mu;l and mix into second tube<br>
+
<div align="left" style="margin-left:30px; margin-right:50px">
-
4. Repeat for the number of dilutions you wish to do (8 should be more than enough)<sup>[2]</sup><br>
+
<ol>
-
5. Take 10 &mu;l from each dilution and spot it on to the agar plate<br>
+
<li>Fill each tube in the dilution with 90 &mu;l of LB.</li>
-
6. Allow droplet to dry and incubate<br>
+
<li>Add 10 &mu;l of the sample to the first tube and mix.</li>
-
<br>
+
<li>From the first tube, remove 10 &mu;l and mix into second tube.</li>
-
The first dilutions will contain a thick lawn of cells and the last dilutions will contain no cells. There should be one drop which contains countable single colonies. From this, you can calculate the number of cells in the original sample. For example, if there 4 colonies on dilution 5, there are 4E4 cells/&mu;l.<br>
+
<li>Repeat for the number of dilutions you wish to do (8 should be more than enough) <sup><span style="color:blue">[1]</span></sup>.</li>
-
<br>
+
<li>Take 10 &mu;l from each dilution and spot it on to the agar plate.</li>
-
<center>
+
<li>Allow droplet to dry and incubate.</li>
-
<B>References<br>
+
</ol>
-
http://openwetware.org/wiki/Cell_counting/plating
+
</div></font></p><br>
 +
<p text-aligne:left style="margin-left:60px; margin-right:60px"><font size="3" color="#F0F8FF" face="Arial regular">
 +
The first dilutions will contain a thick lawn of cells and the last dilutions will contain no cells. There should be one drop which contains countable single colonies. From this, you can calculate the number of cells in the original sample. For example, if there 4 colonies on dilution 5, there are 4E4 cells/&mu;l.</font></p>
 +
 
 +
 
 +
<br><h2><font size="6" color="#F0F8FF" face="Arial regular">References</font></h2>
 +
<font size="3" color="#F0F8FF" face="Arial regular">
 +
<ol>
 +
<li style="margin-left:15px; margin-right:50px; text-align:justify">http://openwetware.org/wiki/Bacterial_cell_culture</li>
 +
</ol>
 +
</font>

Latest revision as of 00:30, 5 October 2013







Lab book | Materials | Protocols

Cell counting/plating

Materials


Procedure

  1. Fill each tube in the dilution with 90 μl of LB.
  2. Add 10 μl of the sample to the first tube and mix.
  3. From the first tube, remove 10 μl and mix into second tube.
  4. Repeat for the number of dilutions you wish to do (8 should be more than enough) [1].
  5. Take 10 μl from each dilution and spot it on to the agar plate.
  6. Allow droplet to dry and incubate.


The first dilutions will contain a thick lawn of cells and the last dilutions will contain no cells. There should be one drop which contains countable single colonies. From this, you can calculate the number of cells in the original sample. For example, if there 4 colonies on dilution 5, there are 4E4 cells/μl.


References

  1. http://openwetware.org/wiki/Bacterial_cell_culture

Retrieved from "http://2013.igem.org/Team:TU_Darmstadt/protocols/Cell_counting/plating"