Team:Kyoto/Protocol

From 2011.igem.org

(Difference between revisions)
(→Protocol)
 
(87 intermediate revisions not shown)
Line 1: Line 1:
-
{{Kyoto_Foreground}}
+
{{Kyoto_Foreground|active_page=method}}
{{Kyoto_Background}}
{{Kyoto_Background}}
{{Kyoto_WikiDesign}}
{{Kyoto_WikiDesign}}
-
<!-- main -->
+
='''Protocol'''=
-
<div id="main">
+
Listed below our protocol for this project.<br>
 +
We hope you find them useful!!
-
= '''Protocol''' =
+
<html><a name="cloning"></a></html>
-
=== Medium for drosophila ===
+
[[File:Cloning.PNG|link=https://2011.igem.org/Team:Kyoto/Cloning|Cloning]]<br>
-
::{| class="wikitable"
+
[[File:Medium.PNG|link=https://2011.igem.org/Team:Kyoto/Medium|Medium]]<br>
-
|+ [http://www.biol.se.tmu.ac.jp/fly/www/standard-medium.html Medium for drosophila]
+
[[File:Measurement.PNG|link=https://2011.igem.org/Team:Kyoto/Measurement|Measurement]]
-
! width="300" |Materials
+
-
! width="300" |Methods
+
-
|-
+
-
|
+
-
* water : 500 mL
+
-
* dry yeast : 20 g
+
-
* corn flour : 45 g
+
-
* glucose : 50 g
+
-
* agarose : 3.5~5 g
+
-
* propionic acid : 1.5 mL
+
-
* 10 % p-hydroxybenzoate in 70 % Eternol : 5 g
+
-
|
+
-
# Stir dry yeast and agarose with about two-thirds of water. Then, autoclave it.
+
-
# Stir corn flour and glucose with the remaining water.
+
-
# Stir 1 and 2, then autoclave it again.
+
-
# after autoclave, add propionic acid and 10 % p-hydroxybenzoate in 70 % Eternol into it.■
+
-
|}
+
-
<br/>
+
-
 
+
-
===M9 medium===
+
-
::{| class="wikitable"
+
-
! width="300" |Materials
+
-
! width="300" |Methods
+
-
|-
+
-
|
+
-
* water : 1 L
+
-
* Na2HPO4: 6 g
+
-
* KH2PO4 : 3 g
+
-
* NaCl : 0.5 g
+
-
* NH4Cl : 1 g
+
-
* 100 mM MgSO4 : 10 ml
+
-
* 20 % glucose : 10 mL
+
-
* 10 mM CaCl2 : 10 ml
+
-
* 100 mM thiamine-HCl : 10 ml
+
-
* 20 % casamino acid : 10 ml
+
-
|
+
-
# Stir Na2HPO4, KH2PO4, NaCl and NH4Cl with water.
+
-
# After autoclave, add 10 ml filter sterilized 100 mM MgSO4, 20 % glucose, 10 mM CaCl2, 100 mM thiamine-HCl.
+
-
# If you need, add 10 ml filter sterilized 20 % casamino acid.
+
-
|}
+
-
<br/>
+
-
 
+
-
===LB medium===
+
-
::{| class="wikitable"
+
-
! width="300" |Materials
+
-
! width="300" |Methods
+
-
|-
+
-
|
+
-
* water : 1 L
+
-
* Tryptone: 10 g
+
-
* Yeast extract : 5 g
+
-
* NaCl : 10 g
+
-
* Agar : 15g
+
-
|
+
-
# Stir Tryptone, Yeast extract and NaCl with water.
+
-
# If you make LB plates, add agar.
+
-
# Autoclave.
+
-
|}
+
-
<br/>
+
-
 
+
-
===SOB medium===
+
-
::{| class="wikitable"
+
-
! width="300" |Materials
+
-
! width="300" |Methods
+
-
|-
+
-
|
+
-
* water : 100 mL
+
-
* Tryptone: 2 g
+
-
* Yeast extract : 0.5 g
+
-
* 5 M NaCl : 200 ul
+
-
* 3 M KCl : 84 ul
+
-
* 1 M MgSO4 : 1 ml
+
-
* 1 M MgCl2 : 1 ml
+
-
|
+
-
# Stir Tryptone and Yeast extract with water.
+
-
# Add 200 ul 5 M NaCl and 84 ul 3 M KCl.
+
-
# After autoclave, add 1 ml filter sterilized 10 mM MgSO4 and 10 mM MgCl2.
+
-
|}
+
-
<br/>
+
-
 
+
-
===SOC medium===
+
-
::{| class="wikitable"
+
-
! width="300" |Materials
+
-
! width="300" |Methods
+
-
|-
+
-
|
+
-
* SOB : 100 ml
+
-
* 2 M glucose: 1 ml
+
-
|
+
-
# Add 2 M glucose to 100 ml SOB.
+
-
|}
+
-
<br/>
+
-
 
+
-
===Buffer TB===
+
-
::{| class="wikitable"
+
-
! width="300" |Materials
+
-
! width="300" |Methods
+
-
|-
+
-
|
+
-
* water : 200 ml
+
-
* PIPES : 3 g
+
-
* CaCl2・2H2O : 0.22 g
+
-
* 3 M KCl : 8.315 ml
+
-
* KOH
+
-
* MnCl2・4H2O : 1.09 g
+
-
|
+
-
# Add 2 M glucose to 100 ml SOB.
+
-
|}
+
-
 
+
-
</div>
+
-
<!-- end of main -->
+

Latest revision as of 02:26, 6 October 2011

Protocol

Listed below our protocol for this project.
We hope you find them useful!!

Cloning
Medium
Measurement

Retrieved from "http://2011.igem.org/Team:Kyoto/Protocol"