Team:UNIPV-Pavia/Material Methods

From 2010.igem.org

(Difference between revisions)
(New page: <!-- originale della home disponibile a https://2010.igem.org/Team:UNIPV-Pavia/homebackup --> <table width="100%" border="0"> <tr> <td colspan="2"> {{UNIPV-Pavia/header}} </td> ...)
 
(28 intermediate revisions not shown)
Line 1: Line 1:
-
<!-- originale della home disponibile a https://2010.igem.org/Team:UNIPV-Pavia/homebackup -->
+
<table width="100%" border="0" >
-
 
+
-
 
+
-
<table width="100%" border="0">
+
<tr>
<tr>
-
      <td colspan="2"> {{UNIPV-Pavia/header}} </td> 
+
    <td colspan="2">{{UNIPV-Pavia/header}}</td>
-
        <!-- <td colspan="2" align="center"><html><img src="http://img337.imageshack.us/img337/7634/logoigem.jpg"  width="100%" /></html></td> -->  
+
</tr>
</tr>
-
 
+
<tr><td align="left" valign="top" width="15%">{{UNIPV-Pavia/menu}}</td><td style="padding:20px">
-
<!-- <tr><td colspan="2"><html><marquee>countdown</marquee></html></td></tr> -->
+
<p align="center" valign="top"><br>
-
 
+
<font size="6"  align="center" valign="top">'''''Material & Methods'''''</font>
-
<tr><td align="left" valign="top" width="20%">{{UNIPV-Pavia/menu}}</td>
+
</p>
-
 
+
<hr>
-
<!-- Contenuti -->
+
-
 
+
-
<td valign="top" >
+
-
 
+
-
=Media & Antibiotics=
+
-
 
+
-
==LB==
+
-
* Add:
+
-
** 10 g/L NaCl
+
-
** 10 g/L Bacto-Tryptone
+
-
** 0.5 g/L Bacto-Yeast Extract
+
-
** ddH2O
+
-
to a sterile pyrex bottle
+
-
* autoclave
+
-
* (add antibiotic when it reaches ~45°C)
+
-
* store at +4°C
+
-
 
+
<br>
<br>
-
==LB Agar==
+
<table valign="center" border="0" width="100%">
-
* Add:
+
<tr><td width="33%" align="center">
-
** 10 g/L NaCl
+
<html><a href="https://2010.igem.org/Team:UNIPV-Pavia/Material_Methods/Protocols">
-
** 10 g/L Bacto-Tryptone
+
<img src="https://static.igem.org/mediawiki/2010/0/03/UNIPV_Pavia_Protocolli.jpg" width="200px" height="200px"/></a>
-
** 0.5 g/L Bacto-Yeast Extract
+
</html></td>
-
** 15 g/L Bacto-Agar
+
    <td width="33%" align="center"><html><a href="https://2010.igem.org/Team:UNIPV-Pavia/Material_Methods/Instruments">
-
** ddH2O
+
<img src="https://static.igem.org/mediawiki/2010/c/c3/UNIPV_Pavia_strumentazione.jpg" width="200px" height="200"/></a>
-
to a sterile 1L flask
+
</html></td>
-
* autoclave
+
    <td width="33%" align="center"><html><a href="https://2010.igem.org/Team:UNIPV-Pavia/Material_Methods/Measurements/Tecan">
-
* (add antibiotic when it reaches ~45°C, shake gently to avoid bubbles)
+
<img src="https://static.igem.org/mediawiki/2010/3/38/UNIPV_Pavia_provette.jpg" width="200px" height="200px"/></a>
-
* pour into Petri plates
+
</html></td>
-
* let them polymerize for ~2-3h
+
</tr>
-
* invert plates and wrap them with aluminium foil and store at +4°C
+
<tr><td width="33%" align="center"><font size="3">[[Team:UNIPV-Pavia/Material_Methods/Protocols|PROTOCOLS]]</font></td>
-
 
+
    <td width="33%" align="center"><font size="3">[[Team:UNIPV-Pavia/Material_Methods/Instruments|INSTRUMENTS]]</font></td>
-
<br>
+
    <td width="33%" align="center"><font size="3">[[Team:UNIPV-Pavia/Material_Methods/Measurements/Tecan|TECAN MEASUREMENTS]]</font></td>
-
==SOB==
+
</tr>
-
* Add:
+
</table>
-
** 5 g/L Bacto-Yeast Extract
+
</td>
-
** 20 g/L Bacto-Tryptone
+
</tr>
-
** 10mM NaCl
+
-
** 2.5mM KCl
+
-
** 10mM MgSO4
+
-
** 10mM MgCl2
+
-
to a sterile pyrex bottle
+
-
* (optional: check that pH is ~6.8, otherwise adjust with NaOH)
+
-
* autoclave
+
-
* (add antibiotic when it reaches ~45°C)
+
-
* store at +4°C
+
-
 
+
-
<br>
+
-
==SOC==
+
-
* SOB+20mM (3.6 g/L) of glucose (add filter-sterilized (0.2um) glucose to autoclaved SOB).
+
-
 
+
-
<br>
+
-
==M9 supplemented with glycerol (M9gly)==
+
-
 
+
-
For 1L of medium, add:
+
-
* 716 ml of autoclaved (and cooled to Tamb) ddH2O
+
-
* 200 ul of autoclaved or filtered (0.2um) CaCl2 0.5 M
+
-
* 200 ml of autoclaved M9 salts 5x (dissolve 56.4 g in 1 liter ddH2O = 5x stock)
+
-
* 34 ml of filtered (0.2um) thiamine hydrochloride MW=337.27g/mol (340 mg in 34 ml)
+
-
* 20 ml of autoclaved MgSO4 0.1 M
+
-
* 20 ml of 10% autoclaved casamino acids (dissolve 50 g in 500 ml = 10% stock)
+
-
* 10 ml of autoclaved 40% glycerol as carbon source
+
-
* mix all the solutions in sterility (each solution must be completely dissolved!)
+
-
* (add antibiotic)
+
-
* store at +4°C, protected from light
+
-
 
+
-
NOTE:
+
-
* M9 salts 5x
+
-
* 10% casamino acids
+
-
can be stored at +4°C
+
-
* MgSO4 0.1 M
+
-
* CaCl2 0.5 M
+
-
* glycerol 40%
+
-
can be stored at room temperature or +4°C
+
-
* thiamine hydrochloride (LIGHT SENSITIVE) is one-shot and must be prepared each time
+
-
 
+
-
 
+
-
 
+
-
</td></tr>
+
</table>
</table>
 +
<br><br>

Latest revision as of 15:00, 22 October 2010
Home
The Project
Materials & Methods
Parts
The Team
Notebook
Biological Safety
Pavia
Gallery
Sponsors


Material & Methods


PROTOCOLS INSTRUMENTS TECAN MEASUREMENTS



Retrieved from "http://2010.igem.org/Team:UNIPV-Pavia/Material_Methods"