Difference between revisions of "Part:BBa K1431812:Experience"

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=Applications of BBa_K1431812=
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After we constructed this part, we tranformed into BL21 & DH5α to test expression effeciency. In the meantime, we did a comparison between BBa_K1431812 and BBa_K1431832 whose promoter is BBa_J23106, a relative weak promoter.
  
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__NOTOC__
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==LB agar plate growth comparison==
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
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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.
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how you used this part and how it worked out.
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===Applications of BBa_K1431812===
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After we constructed this part, it was tranformed into BL21 & DH5α to test expression effeciency. In then meanwhile, we did a comparison between BBa_K1431812 and of which part promoter is changed to BBa_J23106(weak).
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We spread plate at 10:00 on 15th, Oct. Following are pictures which was taken every several hours after placing the plates into a incubator.
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JA is J23100,JB is J23106.
 
JA is J23100,JB is J23106.
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At 10:00 (16th,Oct.)
 
At 10:00 (16th,Oct.)
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<center>https://static.igem.org/mediawiki/parts/a/a8/SUSTC-Shenzhen_Characterization_K09_10.png</center>
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<center>https://static.igem.org/mediawiki/parts/e/e5/SUSTC-Shenzhen_Characterization_K09_10.png</center>
  
  
 
At 11:20 (16th,Oct.)
 
At 11:20 (16th,Oct.)
 
<center>https://static.igem.org/mediawiki/parts/e/e8/SUSTC-Shenzhen_Characterization_K09_11_20.png</center>
 
<center>https://static.igem.org/mediawiki/parts/e/e8/SUSTC-Shenzhen_Characterization_K09_11_20.png</center>
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==LB broth growth comparison==
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We isolated single colonies from the plates and incubate in 2ml LB broth with 34ug/ml chloramphenicol.
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At 00:50 (17th,Oct.)
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<center>https://static.igem.org/mediawiki/parts/e/ed/SUSTC-Shenzhen_K09_L-1.jpg</center>
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At 02:45 (17th,Oct.)
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<center> https://static.igem.org/mediawiki/parts/4/4b/SUSTC-Shenzhen_K09_L-2.jpg</center>
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At 04:55 (17th,Oct.)
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<center> https://static.igem.org/mediawiki/parts/5/5c/SUSTC-Shenzhen_K09_L-3.jpg</center>
  
  
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From observation, expression in BL21 is stronger than in DH5α, and the part only differs in promoter with J23106 unexpected is stronger.
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At 10:40 (17th,Oct.)
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<center> https://static.igem.org/mediawiki/parts/6/69/SUSTC-Shenzhen_K09_L-4.jpg</center>
  
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===User Reviews===
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==Conclusion==
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<!-- DON'T DELETE --><partinfo>BBa_K1431812 StartReviews</partinfo>
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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.
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<partinfo>BBa_K1431812 AddReview number</partinfo>
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<I>Username</I>
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Enter the review inofrmation here.
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<!-- DON'T DELETE --><partinfo>BBa_K1431812 EndReviews</partinfo>
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Latest revision as of 13:05, 23 October 2014

Applications of BBa_K1431812

After we constructed this part, we tranformed into BL21 & DH5α to test expression effeciency. In the meantime, we did a comparison between BBa_K1431812 and BBa_K1431832 whose promoter is BBa_J23106, a relative weak 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 10:40 (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.