Team:Chiba/Notebook/Calendar/3 September 2009

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== Making Fluorescent Transfer Curve for the Practice of Lab Work (Day 2)==
 +
*main culture
 +
10:30  We took 30 mL of the liquid medium that we had made yesterday to conical flask and added prior culture solution 300 &mu;L, then started shake culture at 37 degrees Celsius.
 +
 +
*Making glycerol stock
 +
We kept mixture of 300 &mu;L of 50% glycerol and 300 &mu;L of solution which is cultured yesterday in deep freezer.
 +
 +
 +
*Making Transfer Curves
 +
16:00
 +
 +
We put the solution that has completed main culture in 48 deep wells.
 +
 +
17:15
 +
 +
We shook 48 deep wells at 30 degrees Celsius.
 +
 +
While shaking, we measured OD and GFP fluorescence intensity of sample.
 +
 +
 +
OD
 +
:<table width="500" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
:<td width="100">0 &mu;M AHL</td>
 +
:<td width="100">0.597</td>
 +
:<td width="100">0.590</td>
 +
:<td width="100">0.589</td>
 +
:<td width="100">0.043(control)</td>
 +
:</tr>
 +
:<tr>
 +
:<td>100 &mu;M AHL</td>
 +
:<td>0.592</td>
 +
:<td>0.595</td>
 +
:<td>0.591</td>
 +
:</tr>
 +
:</table>
 +
 +
 +
GFP Fluorescence Intensity
 +
:<table width="500" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
:<td width="100">0 &mu;M AHL</td>
 +
:<td width="100">2.218</td>
 +
:<td width="100">2.043</td>
 +
:<td width="100">2.099</td>
 +
:<td width="100">1.748(control)</td>
 +
:</tr>
 +
:<tr>
 +
:<td width="100">100 &mu;M AHL</td>
 +
:<td>2.139</td>
 +
:<td>2.114</td>
 +
:<td>2.121</td>
 +
:</tr>
 +
:</table>
 +
 +
 +
 +
18:30
 +
 +
We put the sample on plate and measured OD and GFP fluorescence intensity.
 +
 +
OD
 +
:<table width="400" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
:<td width="100">0.624</td>
 +
:<td width="100">0.635</td>
 +
:<td width="100">0.613</td>
 +
:<td width="100">0.040</td>
 +
:</tr>
 +
:<tr>
 +
:<td>0.631</td>
 +
:<td>0.617</td>
 +
:<td>0.601</td>
 +
:</tr>
 +
:</table>
 +
 +
 +
GFP Fluorescence Intensity
 +
:<table width="400" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
:<td width="100">2.119</td>
 +
:<td width="100">2.035</td>
 +
:<td width="100">2.038</td>
 +
:<td width="100">1.718</td>
 +
:</tr>
 +
:<tr>
 +
:<td>2.044</td>
 +
:<td>2.183</td>
 +
:<td>2.105</td>
 +
:</tr>
 +
:</table>
 +
 +
 +
 +
19:24
 +
 +
In a similar way.
 +
 +
OD
 +
:<table width="400" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
:<td width="100">0.741</td>
 +
:<td width="100">0.724</td>
 +
:<td width="100">0.717</td>
 +
:<td width="100">0.040</td>
 +
:</tr>
 +
:<tr>
 +
:<td>0.718</td>
 +
:<td>0.698</td>
 +
:<td>0.678</td>
 +
:</tr>
 +
:</table>
 +
 +
 +
GFP Fluorescence Intensity
 +
:<table width="400" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
:<td width="100">2.071</td>
 +
:<td width="100">2.050</td>
 +
:<td width="100">2.102</td>
 +
:<td width="100">1.666</td>
 +
:</tr>
 +
:<tr>
 +
:<td>2.449</td>
 +
:<td>2.263</td>
 +
:<td>2.344</td>
 +
:</tr>
 +
:</table>
 +
 +
 +
 +
20:23
 +
 +
In a similar way.
 +
 +
OD
 +
:<table width="400" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
:<td width="100">0.774</td>
 +
:<td width="100">0.742</td>
 +
:<td width="100">0.742</td>
 +
:<td width="100">0.040</td>
 +
:</tr>
 +
:<tr>
 +
:<td>0.735</td>
 +
:<td>0.723</td>
 +
:<td>0.760</td>
 +
:</tr>
 +
:</table>
 +
 +
 +
GFP Fluorescence Intensity
 +
:<table width="400" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
:<td width="100">2.158</td>
 +
:<td width="100">2.227</td>
 +
:<td width="100">2.364</td>
 +
:<td width="100">1.646</td>
 +
:</tr>
 +
:<tr>
 +
:<td>2.815</td>
 +
:<td>2.767</td>
 +
:<td>2.756</td>
 +
:</tr>
 +
:</table>
-
*本培養/main culture
 
-
10:30 昨日作成したAmp-Cm液体培地30 mLとプレ培養した溶液300 μLを三角フラスコにとり、37℃で振とうした。/we took the liquid medium that we had made yesterday to conical flask 30 mL and added prior culture 300 uL, then started shake culture at 37 degrees Celsius.
 
-
*グリスト作成/Making glycerol stock
+
21:18
-
50%グリセロール300 μLとプレ培養した溶液300 μLを1.5 mLマイクロチューブにとり、Deep Freezerに保存した。/We kept mixture of 300 μL of 50% glycerol and 300 μL of solution which is cultured yesterday in deep freezer.
+
 +
In a similar way.
-
*トランスファーカーブ作成実験/Making transfer curves
+
OD
-
16:00 本培養が終わった溶液を10倍希釈し、48穴Deep wellに入れた。/We put the solution that has completed main culture in 48 deep wells.
+
:<table width="400" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
:<td width="100">0.861</td>
 +
:<td width="100">0.813</td>
 +
:<td width="100">0.825</td>
 +
:<td width="100">0.039</td>
 +
:</tr>
 +
:<tr>
 +
:<td>0.755</td>
 +
:<td>0.818</td>
 +
:<td>0.849</td>
 +
:</tr>
 +
:</table>
-
17:15 48穴Deep wellを30℃で振とうした。/We shook 48 deep wells at 30 degrees Celsius.
 
-
振とうしている間にT=0の時点での試料のOD値および蛍光強度を測定した。/While shaking, we measured OD and GFP fluorescence intensity of sample.
+
GFP Fluorescence Intensity
 +
:<table width="400" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
:<td width="100">2.062</td>
 +
:<td width="100">2.775</td>
 +
:<td width="100">2.673</td>
 +
:<td width="100">1.814</td>
 +
:</tr>
 +
:<tr>
 +
:<td>3.615</td>
 +
:<td>3.510</td>
 +
:<td>3.453</td>
 +
:</tr>
 +
:</table>
-
18:15 プレートに試料をとり、OD値と蛍光強度を測定した。
 
-
19:15 同様
 
-
20:15 同様
+
22:56
 +
'''We stopped this experiment.'''
-
[[Image:Chi_20090903_1.JPG|200px]]
+
== Handwritten Lab Notebook ==
-
※写真は本培養の開始までです。
+
[[Image:Chi_20090903_1.JPG|200px]] [[Image:Chiba Labwork 03Sep09.jpg|200px]] [[Image:Chiba Labwork 03Sep09-2.jpg|200px]]

Latest revision as of 17:44, 23 September 2009

>Go to the Notebook page

(2_September_2009 <|>4_September_2009)


Making Fluorescent Transfer Curve for the Practice of Lab Work (Day 2)

  • main culture

10:30 We took 30 mL of the liquid medium that we had made yesterday to conical flask and added prior culture solution 300 μL, then started shake culture at 37 degrees Celsius.

  • Making glycerol stock

We kept mixture of 300 μL of 50% glycerol and 300 μL of solution which is cultured yesterday in deep freezer.


  • Making Transfer Curves

16:00

We put the solution that has completed main culture in 48 deep wells.

17:15

We shook 48 deep wells at 30 degrees Celsius.

While shaking, we measured OD and GFP fluorescence intensity of sample.


OD

0 μM AHL 0.597 0.590 0.589 0.043(control)
100 μM AHL 0.592 0.595 0.591


GFP Fluorescence Intensity

0 μM AHL 2.218 2.043 2.099 1.748(control)
100 μM AHL 2.139 2.114 2.121


18:30

We put the sample on plate and measured OD and GFP fluorescence intensity.

OD

0.624 0.635 0.613 0.040
0.631 0.617 0.601


GFP Fluorescence Intensity

2.119 2.035 2.038 1.718
2.044 2.183 2.105


19:24

In a similar way.

OD

0.741 0.724 0.717 0.040
0.718 0.698 0.678


GFP Fluorescence Intensity

2.071 2.050 2.102 1.666
2.449 2.263 2.344


20:23

In a similar way.

OD

0.774 0.742 0.742 0.040
0.735 0.723 0.760


GFP Fluorescence Intensity

2.158 2.227 2.364 1.646
2.815 2.767 2.756


21:18

In a similar way.

OD

0.861 0.813 0.825 0.039
0.755 0.818 0.849


GFP Fluorescence Intensity

2.062 2.775 2.673 1.814
3.615 3.510 3.453


22:56

We stopped this experiment.


Handwritten Lab Notebook

Chi 20090903 1.JPG Chiba Labwork 03Sep09.jpg Chiba Labwork 03Sep09-2.jpg

Retrieved from "http://2009.igem.org/Team:Chiba/Notebook/Calendar/3_September_2009"