Difference between revisions of "Team:Nagahama/Protocol"

 
Line 553: Line 553:
 
<br/>
 
<br/>
 
Protocols
 
Protocols
 +
<Br>
 
Our Lab's Protocols  
 
Our Lab's Protocols  
 +
<Br>
 
Medium
 
Medium
 +
<Br>
 
LB medium (100 mL liquid)
 
LB medium (100 mL liquid)
 +
<Br>
 
1.Measure 1g Tripton  
 
1.Measure 1g Tripton  
 +
<Br>
 
2.Measure 0.5g Yeast Extract  
 
2.Measure 0.5g Yeast Extract  
 +
<Br>
 
3.Measure 1g Nacl  
 
3.Measure 1g Nacl  
 +
<Br>
 
4.Add 100mL H2O  
 
4.Add 100mL H2O  
 +
<Br>
 
5.autoclave(121℃ 20min)  
 
5.autoclave(121℃ 20min)  
 +
<Br>
 
2×YT medium (100mL liquid)
 
2×YT medium (100mL liquid)
 +
<Br>
 
1.Measure 1.6g Tripton  
 
1.Measure 1.6g Tripton  
 +
<Br>
 
2.Measure 1g Yeast Extract  
 
2.Measure 1g Yeast Extract  
 +
<Br>
 
3.Measure 0.5g Nacl  
 
3.Measure 0.5g Nacl  
 +
<Br>
 
4.Add 100mL H2O  
 
4.Add 100mL H2O  
 +
<Br>
 
5.autoclave(121℃ 20min)  
 
5.autoclave(121℃ 20min)  
 
+
<Br>
 +
<Br>
 
DNA work
 
DNA work
 +
<Br>
 
Agarose gel(100mL)
 
Agarose gel(100mL)
 +
<Br>
 
Method of Making 0.7% Agarose gel  
 
Method of Making 0.7% Agarose gel  
 +
<Br>
 
1.Measure 0.7g Agarose  
 
1.Measure 0.7g Agarose  
 +
<Br>
 
2.Add 100mL TAE buffer  
 
2.Add 100mL TAE buffer  
3.Heat(till agarose melted)*We used a microwave oven.  
+
<Br>
 +
3.Heat(till agarose melted)*We used a microwave oven.
 +
<Br>
 
4.Pur agarose into a gel maker  
 
4.Pur agarose into a gel maker  
 +
<Br>
 
5.Set a comb  
 
5.Set a comb  
 +
<Br>
 
6.Wait till agarose curdles  
 
6.Wait till agarose curdles  
 
+
<Br>
 +
<Br>
 
isoprenoid production
 
isoprenoid production
 +
<Br>
 
Because of its high hydrophobicity and low volatility, decane was chosen to extract and solubilize farnesol from the culture broth. The decane was overlaidy in the two-phase culture media, but it did not affect the cell growth, and farnesol could be well solubilized in the decane phase with negligible volatile loss. We adopt used 1 mL of decane to overlayid to the 5 mL of culture broth. Two-phase The culture of E. coli JM109 (BBa_K165025) was carried out in 2YT medium containing 1% glycerol at 29°C for 48 h. The decane phase of the two-phase culture was collected to analyze the farnesol content by GC-MS.
 
Because of its high hydrophobicity and low volatility, decane was chosen to extract and solubilize farnesol from the culture broth. The decane was overlaidy in the two-phase culture media, but it did not affect the cell growth, and farnesol could be well solubilized in the decane phase with negligible volatile loss. We adopt used 1 mL of decane to overlayid to the 5 mL of culture broth. Two-phase The culture of E. coli JM109 (BBa_K165025) was carried out in 2YT medium containing 1% glycerol at 29°C for 48 h. The decane phase of the two-phase culture was collected to analyze the farnesol content by GC-MS.
 +
<Br>

Latest revision as of 23:20, 19 October 2016

Medium

LB medium (100 mL liquid) 1.Measure 1g Tripton 2.Measure 0.5g Yeast Extract 3.Measure 1g Nacl 4.Add 100mL H2O 5.autoclave(121℃ 20min)


2×YT medium (100mL liquid) 1.Measure 1.6g Tripton 2.Measure 1g Yeast Extract 3.Measure 0.5g Nacl 4.Add 100mL H2O 5.autoclave(121℃ 20min)
Protocols
Our Lab's Protocols
Medium
LB medium (100 mL liquid)
1.Measure 1g Tripton
2.Measure 0.5g Yeast Extract
3.Measure 1g Nacl
4.Add 100mL H2O
5.autoclave(121℃ 20min)
2×YT medium (100mL liquid)
1.Measure 1.6g Tripton
2.Measure 1g Yeast Extract
3.Measure 0.5g Nacl
4.Add 100mL H2O
5.autoclave(121℃ 20min)

DNA work
Agarose gel(100mL)
Method of Making 0.7% Agarose gel
1.Measure 0.7g Agarose
2.Add 100mL TAE buffer
3.Heat(till agarose melted)*We used a microwave oven.
4.Pur agarose into a gel maker
5.Set a comb
6.Wait till agarose curdles

isoprenoid production
Because of its high hydrophobicity and low volatility, decane was chosen to extract and solubilize farnesol from the culture broth. The decane was overlaidy in the two-phase culture media, but it did not affect the cell growth, and farnesol could be well solubilized in the decane phase with negligible volatile loss. We adopt used 1 mL of decane to overlayid to the 5 mL of culture broth. Two-phase The culture of E. coli JM109 (BBa_K165025) was carried out in 2YT medium containing 1% glycerol at 29°C for 48 h. The decane phase of the two-phase culture was collected to analyze the farnesol content by GC-MS.