Difference between revisions of "Part:BBa K649202:Experience"

 
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
−
This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
+
 
−
how you used this part and how it worked out.
+
  
 
===Applications of BBa_K649202===
 
===Applications of BBa_K649202===
 +
 +
'''[Sample]'''<br / >
 +
<table border="1">
 +
<tr>
 +
  <th colspan="2">sample</th>
 +
  <th>arabinose</th>
 +
</tr>
 +
<tr>
 +
  <td>1</td>
 +
  <td>PlacIQ-lox71-RFP-lox66-GFP(pSB3K3)<br />PBAD/araC-Cre(pSB1A2)</td>
 +
  <td>+</td>
 +
</tr>
 +
<tr>
 +
  <td>2</td>
 +
  <td>PlacIQ-lox71-RFP-lox66-GFP(pSB3K3)<br />PBAD/araC-Cre(pSB1A2)</td>
 +
  <td>-</td>
 +
</tr>
 +
<tr>
 +
  <td>3</td>
 +
  <td>PlacIQ-lox71-RFP-lox66-GFP(pSB3K3)<br /> : negative control </td>
 +
  <td>+</td>
 +
</tr>
 +
</table>
 +
 +
 +
<br / >
 +
<br / >
 +
 +
'''[Method]'''<br / >
 +
1. Strain JM2.300 with both Pbad/araC-Cre and PlacIQ-lox2272-RFP-lox2272-GFP in it was cultured at 37℃ in 3ml of LB medium containing ampicillin (6 µL) and kanamycin(3.6 µL). Same kind of strain having only PlacIQ-lox2272-RFP-lox2272-GFP was cultured in same volume of LB containing kanamycin (3.6 µL) and carbenicillin(4 µL). These were cultured until OD 1.6.
 +
<br / ><br / ><br / >
 +
2. Each cultured medium was 6 times diluted in the medium and three samples (3 ml each)  were dispensed from those. Among them, two were induced by arabinose (2 M, 75 µL).
 +
<br / ><br / ><br / >
 +
3. After 30 min from induction, 1000 times dilution and 100000 times dilution of each sample were plated and incubated in 37°C about 12 hours. The florescence of plates was examined by FLC and it was taken picture. About the rest of the samples, strains were harvested by centrifugation and suspended by adding 1 mL of PBS (phosphate-buffered saline). The last OD of PBS solution was approximately 0.4. We dispensed 700 µL of each suspension into a disposable tube through a cell strainer, and fluorescence intensity of each cell was measured with a flow cytometer of Becton, Dickinson and Company.
 +
<br / ><br / ><br / >
 +
4. In the same way as 3, 9 more samples were examined at period 1hr, 2hr, and 4hr.
  
 
===User Reviews===
 
===User Reviews===

Revision as of 22:42, 3 October 2011


Applications of BBa_K649202

[Sample]

sample arabinose
1 PlacIQ-lox71-RFP-lox66-GFP(pSB3K3)
PBAD/araC-Cre(pSB1A2)		 +
2 PlacIQ-lox71-RFP-lox66-GFP(pSB3K3)
PBAD/araC-Cre(pSB1A2)		 -
3 PlacIQ-lox71-RFP-lox66-GFP(pSB3K3)
 : negative control 		+




[Method]
1. Strain JM2.300 with both Pbad/araC-Cre and PlacIQ-lox2272-RFP-lox2272-GFP in it was cultured at 37℃ in 3ml of LB medium containing ampicillin (6 µL) and kanamycin(3.6 µL). Same kind of strain having only PlacIQ-lox2272-RFP-lox2272-GFP was cultured in same volume of LB containing kanamycin (3.6 µL) and carbenicillin(4 µL). These were cultured until OD 1.6.


2. Each cultured medium was 6 times diluted in the medium and three samples (3 ml each) were dispensed from those. Among them, two were induced by arabinose (2 M, 75 µL).


3. After 30 min from induction, 1000 times dilution and 100000 times dilution of each sample were plated and incubated in 37°C about 12 hours. The florescence of plates was examined by FLC and it was taken picture. About the rest of the samples, strains were harvested by centrifugation and suspended by adding 1 mL of PBS (phosphate-buffered saline). The last OD of PBS solution was approximately 0.4. We dispensed 700 µL of each suspension into a disposable tube through a cell strainer, and fluorescence intensity of each cell was measured with a flow cytometer of Becton, Dickinson and Company.


4. In the same way as 3, 9 more samples were examined at period 1hr, 2hr, and 4hr.

User Reviews

UNIQ4855946112c1f26c-partinfo-00000000-QINU UNIQ4855946112c1f26c-partinfo-00000001-QINU