Team:Nagahama/Protocol
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.
