Difference between revisions of "Part:BBa K4060106:Experience"

(Assembly of PreB3 (pSB1C3))
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Fig. 1 Agarose electrophoresis in order to verify Pre B3 (Construct 3). Lane 1, 100bp ladder. Lane 1’, partial of Life 1 kb Plus ladder. Lane 2,3,4 Pre B3 digested with BamHI and HindIII.
 
Fig. 1 Agarose electrophoresis in order to verify Pre B3 (Construct 3). Lane 1, 100bp ladder. Lane 1’, partial of Life 1 kb Plus ladder. Lane 2,3,4 Pre B3 digested with BamHI and HindIII.
  
====1. The RFP expression of Construct 3====
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====2. The RFP expression of Construct 3====
 
We used RFP(E1010) to monitor the expression of TetR, so that we can figure out how L-arabinose will influence the expression of TetR in our Design 1.
 
We used RFP(E1010) to monitor the expression of TetR, so that we can figure out how L-arabinose will influence the expression of TetR in our Design 1.
  
 
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<img src="https://2021.igem.org/wiki/images/c/c5/T--NYCU-Taipei--PreB3.png" height="475px" width="750px"/>
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<img src="https://2021.igem.org/wiki/images/f/fb/T--NYCU-Taipei--kilksw_result_1.png" height="475px" width="750px"/>
 
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<img src="https://2021.igem.org/wiki/images/c/c5/T--NYCU-Taipei--PreB3.png" height="475px" width="750px"/>
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<img src="https://2021.igem.org/wiki/images/3/34/T--NYCU-Taipei--kilksw_result_2.png" height="475px" width="750px"/>
 
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Latest revision as of 14:31, 7 December 2021


1. Assembly of PreB3 (pSB1C3)

The gene fragment Pre B3 is composed of Pre A1 and Pre A3. We digested the vector fragment Pre A1 with SpeI and PstI and the insert fragment Pre A3 with XbaI and PstI. We ligated these fragments together to form Pre B3.

Fig. 1 Agarose electrophoresis in order to verify Pre B3 (Construct 3). Lane 1, 100bp ladder. Lane 1’, partial of Life 1 kb Plus ladder. Lane 2,3,4 Pre B3 digested with BamHI and HindIII.

2. The RFP expression of Construct 3

We used RFP(E1010) to monitor the expression of TetR, so that we can figure out how L-arabinose will influence the expression of TetR in our Design 1.

Fig. 2 The RFP intensity of construct 3(Pre B3) from 0 hr~ 24hrs in different L-arabinose concentrations.

Fig. 3 After 24 hrs cultivation, the RFP intensity of construct 3 in different L-arabinose concentrations

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