Team:UT-Tokyo/Data/Method

From 2011.igem.org

(Difference between revisions)
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Before use, add 10ml Mg sol.
Before use, add 10ml Mg sol.
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;Mg sol.
+
;:Mg sol.
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:{|
+
::{|
|MgCl<span style="vertical-align: sub;">2</span>
|MgCl<span style="vertical-align: sub;">2</span>
|500mM
|500mM
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<div style="float:right; width:487.5px;">
<div style="float:right; width:487.5px;">
*50&times; TAE
*50&times; TAE
 +
{|
 +
!align="left" style="width:200px"|reagents
 +
!align="right" style="width:80px"|final conc.
 +
!align="right" style="width:80px"|amount
 +
|-
 +
|Tris
 +
|align="right"|2M
 +
|align="right"|242g
 +
|-
 +
|CH<span style="vertical-align: sub;">3</span>COOH
 +
|align="right"|1M
 +
|align="right"|57.1mL
 +
|-
 +
|EDTA (0.5M, pH=8.0)
 +
|align="right"|0.05M
 +
|align="right"|100ml
 +
|-
 +
|style="border-bottom: 2px solid"|MilliQ
 +
|style="text-align:center; border-bottom: 2px solid"|-
 +
|style="text-align:right; border-bottom: 2px solid"|1000ml
 +
|-
 +
|colspan="2"|Total
 +
|style="text-align:right"|1L
 +
|}
 +
*TB
*TB
 +
{|
 +
!align="left" style="width:200px"|reagents
 +
!align="right" style="width:80px"|final conc.
 +
!align="right" style="width:80px"|amount
 +
|-
 +
|KOH 500mM sol.
 +
|align="right"|250mMM
 +
|align="right"|242g
 +
|-
 +
|PIPES 500mM sol.
 +
|align="right"|10mM
 +
|align="right"|2ml
 +
|-
 +
|CaCl<span style="vertical-align: sub;">2</span> 750mM sol.
 +
|align="right"15mM
 +
|align="right"|2ml
 +
|-
 +
|KCl 2.5M sol.
 +
|align="right"|250mM
 +
|align="right"|10ml
 +
|-
 +
|MnCl<span style="vertical-align: sub;">2</span> 550mM sol.
 +
|align="right"|55mM
 +
|align="right"|10ml
 +
|-
 +
|style="border-bottom: 2px solid"|MilliQ
 +
|style="text-align:center; border-bottom: 2px solid"|-
 +
|style="text-align:right; border-bottom: 2px solid"|100ml
 +
|-
 +
|colspan="2"|Total
 +
|style="text-align:right"|100ml
 +
|}
</div>
</div>
<div style="clear:both">
<div style="clear:both">
</div>
</div>
-
 
-
 
==Cultures==
==Cultures==

Revision as of 15:59, 20 September 2011

Reagents

  • LB broth
reagents final conc. amount
Bacto trypton 1% 1g
NaCl 0.5% 0.5g
Yeast extracs 0.5% 0.5g
MilliQ - 100ml
Total 100ml


  • SOB broth
reagents final conc. amount
Bacto trypton 2% 20g
NaCl 0.05% 0.5g
Yeast extracs 0.5% 5g
KCl (250mM) 2.5mM 10ml
MilliQ - 1000ml
Total 1L

Before use, add 10ml Mg sol.

Mg sol.
MgCl2 500mM
MgSO4 500mM


  • M9
  • 50× TAE
reagents final conc. amount
Tris 2M 242g
CH3COOH 1M 57.1mL
EDTA (0.5M, pH=8.0) 0.05M 100ml
MilliQ - 1000ml
Total 1L
  • TB
reagents final conc. amount
KOH 500mM sol. 250mMM 242g
PIPES 500mM sol. 10mM 2ml
CaCl2 750mM sol. align="right"15mM 2ml
KCl 2.5M sol. 250mM 10ml
MnCl2 550mM sol. 55mM 10ml
MilliQ - 100ml
Total 100ml

Cultures

Extraction of DNA from cells

Amplification of DNA

Analysis of DNA

Assays