Team:UT-Tokyo/Data/Method

From 2011.igem.org

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===colony PCR===
===colony PCR===
-
==Amplification of DNA==
+
==Purification of DNA==
 +
 
 +
===Miniprep===
 +
 
 +
===Gel extraction===
 +
 
 +
===PCR clean-up===
 +
 
 +
===Ethanol precipitation===
==Analysis of DNA==
==Analysis of DNA==

Revision as of 02:18, 22 September 2011

Reagents

  • 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.
reagents final conc. amount
MgCl2(H2O)6 1M 20.33g
MgSO4(H2O)7 1M 24.648g
MilliQ - 100ml
Total 100ml


  • 20× M9 medium
reagents final conc. amount
Na2HPO4 - 6.0g
KH2PO4 - 3.0g
NaCL - 0.5g
NH4Cl - 1.0g
MilliQ - 50ml
Total 50ml

After A.C. , add following reagents to 1L M9 medium

reagents final conc. amount
1M MgSO4 - 1.0ml
2M Glucose - 5.6ml
1% Thiamine - 1.0ml
1M CaCl2 - 0.1ml
  • 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


  • 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. 250mM 242g
PIPES 500mM sol. 10mM 2ml
CaCl2 750mM sol. 15mM 2ml
KCl 2.5M sol. 250mM 10ml
MnCl2 550mM sol. 55mM 10ml
MilliQ - 100ml
Total 100ml

Transformation

Making Competent E. coli cell

Transformation of E. coli

Assembly parts

Digest

Ligation

colony PCR

Purification of DNA

Miniprep

Gel extraction

PCR clean-up

Ethanol precipitation

Analysis of DNA

Assays

Retrieved from "http://2011.igem.org/Team:UT-Tokyo/Data/Method"