Difference between revisions of "Part:BBa K1431832:Experience"

 
(→‎Applications of BBa_K1431832)
Line 5: Line 5:
  
 
===Applications of BBa_K1431832===
 
===Applications of BBa_K1431832===
 +
After we constructed this part, we tranformed into BL21 & DH5α to test expression effeciency. In the meantime, we did a comparison between BBa_K1431832 and BBa_K1431812 whose promoter is BBa_J23100, a relative strong promoter.
 +
 +
==LB agar plate growth comparison==
 +
We spread the plate for BL21 at 10:00am on 15th, Oct and DH5α at 11:00am on 15th, Oct. The plate contains 34ug/ml chloramphenicol. Following pictures were taken every several hours after placing the plates into a 37℃ incubator.
 +
 +
JA is J23100,JB is J23106.
 +
 +
At 00:30 (16th,Oct.)
 +
<center>https://static.igem.org/mediawiki/parts/2/2e/SUSTC-Shenzhen_Characterization_K09_0：30.png</center>
 +
 +
 +
At 1:50  (16th,Oct.)
 +
<center>https://static.igem.org/mediawiki/parts/1/19/SUSTC-Shenzhen_Characterization_K09_1_50.png</center>
 +
 +
 +
At 3:00  (16th,Oct.)
 +
<center>https://static.igem.org/mediawiki/parts/d/d7/SUSTC-Shenzhen_Characerization_K09_3.png</center>
 +
 +
 +
At 6:00  (16th,Oct.)
 +
<center>https://static.igem.org/mediawiki/parts/8/83/SUSTC-Shenzhen_Characterization_K09_6.png</center>
 +
 +
 +
At 7:40  (16th,Oct.)
 +
<center>https://static.igem.org/mediawiki/parts/b/b3/SUSTC-Shenzhen_Characterization_K09_7_40.png</center>
 +
 +
 +
At 8:00  (16th,Oct.)
 +
<center>https://static.igem.org/mediawiki/parts/a/a8/SUSTC-Shenzhen_Characterization_K09_9.png</center>
 +
 +
 +
At 10:00 (16th,Oct.)
 +
<center>https://static.igem.org/mediawiki/parts/e/e5/SUSTC-Shenzhen_Characterization_K09_10.png</center>
 +
 +
 +
At 11:20 (16th,Oct.)
 +
<center>https://static.igem.org/mediawiki/parts/e/e8/SUSTC-Shenzhen_Characterization_K09_11_20.png</center>
 +
 +
==LB broth growth comparison==
 +
We isolated single colonies from the plates and incubate in 2ml LB broth with 34ug/ml chloramphenicol.
 +
 +
At 00:50 (17th,Oct.)
 +
<center>https://static.igem.org/mediawiki/parts/e/ed/SUSTC-Shenzhen_K09_L-1.jpg</center>
 +
 +
 +
At 02:45 (17th,Oct.)
 +
<center> https://static.igem.org/mediawiki/parts/4/4b/SUSTC-Shenzhen_K09_L-2.jpg</center>
 +
 +
 +
At 04:55 (17th,Oct.)
 +
<center> https://static.igem.org/mediawiki/parts/5/5c/SUSTC-Shenzhen_K09_L-3.jpg</center>
 +
 +
 +
At 22:40 (17th,Oct.)
 +
<center> https://static.igem.org/mediawiki/parts/6/69/SUSTC-Shenzhen_K09_L-4.jpg</center>
 +
 +
==Conclusion==
 +
From the above observations, we found that expression in BL21 was stronger than that in DH5α, and the strength of promoter BBa_J23100 was unexpectedly weaker than BBa_J23106 provided by the 2014 iGEM kit plate.
  
 
===User Reviews===
 
===User Reviews===

Revision as of 02:19, 18 October 2014


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_K1431832

After we constructed this part, we tranformed into BL21 & DH5α to test expression effeciency. In the meantime, we did a comparison between BBa_K1431832 and BBa_K1431812 whose promoter is BBa_J23100, a relative strong promoter.

LB agar plate growth comparison

We spread the plate for BL21 at 10:00am on 15th, Oct and DH5α at 11:00am on 15th, Oct. The plate contains 34ug/ml chloramphenicol. Following pictures were taken every several hours after placing the plates into a 37℃ incubator.

JA is J23100,JB is J23106.

At 00:30 (16th,Oct.)

https://static.igem.org/mediawiki/parts/2/2e/SUSTC-Shenzhen_Characterization_K09_0：30.png


At 1:50 (16th,Oct.)

SUSTC-Shenzhen_Characterization_K09_1_50.png


At 3:00 (16th,Oct.)

SUSTC-Shenzhen_Characerization_K09_3.png


At 6:00 (16th,Oct.)

SUSTC-Shenzhen_Characterization_K09_6.png


At 7:40 (16th,Oct.)

SUSTC-Shenzhen_Characterization_K09_7_40.png


At 8:00 (16th,Oct.)

SUSTC-Shenzhen_Characterization_K09_9.png


At 10:00 (16th,Oct.)

SUSTC-Shenzhen_Characterization_K09_10.png


At 11:20 (16th,Oct.)

SUSTC-Shenzhen_Characterization_K09_11_20.png

LB broth growth comparison

We isolated single colonies from the plates and incubate in 2ml LB broth with 34ug/ml chloramphenicol.

At 00:50 (17th,Oct.)

SUSTC-Shenzhen_K09_L-1.jpg


At 02:45 (17th,Oct.)

SUSTC-Shenzhen_K09_L-2.jpg


At 04:55 (17th,Oct.)

SUSTC-Shenzhen_K09_L-3.jpg


At 22:40 (17th,Oct.)

SUSTC-Shenzhen_K09_L-4.jpg

Conclusion

From the above observations, we found that expression in BL21 was stronger than that in DH5α, and the strength of promoter BBa_J23100 was unexpectedly weaker than BBa_J23106 provided by the 2014 iGEM kit plate.

User Reviews

UNIQ0e8e441fb0dc3ad4-partinfo-00000000-QINU UNIQ0e8e441fb0dc3ad4-partinfo-00000001-QINU