Difference between revisions of "Part:BBa K4117666:Experience"

 
(→‎Applications of BBa_K4117666)
 
(9 intermediate revisions by the same user not shown)
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
 
This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
Line 5: Line 4:
  
 
===Applications of BBa_K4117666===
 
===Applications of BBa_K4117666===
 +
We added the GFP downstream of the RNA thermometer (RBS) to characterize the efficiency of RNA thermometer.<br>
 +
 +
<center>[[File:T--BNU-China--GFPpathway.png|500px]]</center>
 +
<center>Fig.1 RNA thermometer verification pathway</center><br>
 +
 +
In order to verify the function of RNA thermometers, We cultured the strain at 37°C overnight, and then aliquots, adding pre-prepared flasks filled with 37°C and 28°C LB medium to make the final volume 20ml, then measured the initial OD600 and GFP fluorescence intensity. Samples were taken once every half-hour, OD600 and GFP fluorescence intensity were measured.<br>
 +
 +
GFP measurement results are shown in '''Fig.2'''.<br>
 +
 +
<center>[[File:T--BNU-China--OD600GFP.png|500px]]</center><br>
 +
 +
<center>Fig.2 GFP fluorescence intensity after incubating in different temperatures</center><br>
 +
 +
And we used a fluorescence microscope for observation. The bacterial liquid under the fluorescence microscope is as '''Fig. 3''':<br>
 +
 +
<center>[[File:T--BNU-China--6GFP.png|500px]]</center><br>
 +
 +
<center>Fig.3 Images under fluorescene microscope (fluorescene microscope 10 x 100)</center>
 +
<center> (A) No GFP in engineerd bacteria incubated at 28℃ (B) Bright field image of bacteria incubated at 28℃</center>
 +
<center> (C) The field of view after superposition of A and B (D) GFP in engineerd bacteria incubated at 37℃</center>
 +
<center> (E) Bright field image of bacteria incubated at 37℃ (F) The field of view after superposition of D and E</center><br>
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 08:41, 12 October 2022

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K4117666

We added the GFP downstream of the RNA thermometer (RBS) to characterize the efficiency of RNA thermometer.

T--BNU-China--GFPpathway.png
Fig.1 RNA thermometer verification pathway

In order to verify the function of RNA thermometers, We cultured the strain at 37°C overnight, and then aliquots, adding pre-prepared flasks filled with 37°C and 28°C LB medium to make the final volume 20ml, then measured the initial OD600 and GFP fluorescence intensity. Samples were taken once every half-hour, OD600 and GFP fluorescence intensity were measured.

GFP measurement results are shown in Fig.2.

T--BNU-China--OD600GFP.png

Fig.2 GFP fluorescence intensity after incubating in different temperatures

And we used a fluorescence microscope for observation. The bacterial liquid under the fluorescence microscope is as Fig. 3:

T--BNU-China--6GFP.png

Fig.3 Images under fluorescene microscope (fluorescene microscope 10 x 100)
(A) No GFP in engineerd bacteria incubated at 28℃ (B) Bright field image of bacteria incubated at 28℃
(C) The field of view after superposition of A and B (D) GFP in engineerd bacteria incubated at 37℃
(E) Bright field image of bacteria incubated at 37℃ (F) The field of view after superposition of D and E

User Reviews

UNIQ0b88c49aea8123d3-partinfo-00000000-QINU UNIQ0b88c49aea8123d3-partinfo-00000001-QINU