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Team:UT-Tokyo/Data/Method
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==Reagents== | ==Reagents== | ||
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*SOB broth | *SOB broth | ||
{| | {| | ||
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|align="right"|1.0ml | |align="right"|1.0ml | ||
|- | |- | ||
| - | |1M CaCl<span class="sub">2</ | + | |1M CaCl<span class="sub">2</span> |
|align="right"|- | |align="right"|- | ||
|align="right"|0.1ml | |align="right"|0.1ml | ||
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<div class="float-right"> | <div class="float-right"> | ||
| + | *LB broth | ||
| + | {| | ||
| + | !align="left" style="width:200px"|reagents | ||
| + | !align="right" style="width:80px"|final conc. | ||
| + | !align="right" style="width:80px"|amount | ||
| + | |- | ||
| + | |Bacto trypton | ||
| + | |align="right"|1% | ||
| + | |align="right"|1g | ||
| + | |- | ||
| + | |NaCl | ||
| + | |align="right"|0.5% | ||
| + | |align="right"|0.5g | ||
| + | |- | ||
| + | |Yeast extracs | ||
| + | |align="right"|0.5% | ||
| + | |align="right"|0.5g | ||
| + | |- | ||
| + | |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 | ||
| + | |} | ||
| + | |||
*50× TAE | *50× TAE | ||
{| | {| | ||
Revision as of 12:37, 21 September 2011
Method
iGEM UT-Tokyo
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 | |
Cultures
Extraction of DNA from cells
Amplification of DNA
Analysis of DNA
Assays
