![](https://parts.igem.org/images/partbypart/icon_generator.png)
Generator
HIC genera
Part:BBa_K1602020
Designed by: Sebastian Barthel, Steven Pilger, Jonas Sindlinger, Bianca Reisinger, Sven Jager Group: iGEM15_TU_Darmstadt (2015-09-16)
Inducible generator of Humicola insolens cutinase
This generator overexpress Humicola insolens cutinase (HIC) by using a T7 promoter system. For more information about the Humicola insolens cutinase (HIC) have a look on the coding sequence.
Results
Final volume of the plate was 200µl containig:
- 142µl Buffer (Na2HPO4 pH 7.0)
- bromothymol blue 10% (v/v)
- TES protein fraction HiC (ranging from 5µl to 30µl; concentration unknown)
- 8µl prepolymer (dissolved in Triton100 and DMSO 1:50)
- TES protein fraction (from BL21 cells ranging from 5µl to 30µl; as a control)
The negative control was done by just adding buffer to the well.
The 96 well microplate was loaded as depicted in the picture below: </p>
![](https://static.igem.org/mediawiki/2015/thumb/0/06/Da15_hic_plate.png/800px-Da15_hic_plate.png)
The assay was run in a in a TECAN® Infinite 200 PRO multi plate reader for 100 kinetic cycles, each 5 mins long and with 25 photo pulses per cycle. The reader was heated to the appropriate temperature of 42° celsius. Absorbance was measured at the absorption maximum of BTB which in this case is 620nm.
Pictures of the multi plate
![](https://static.igem.org/mediawiki/2015/thumb/8/81/Da15_1_hic.jpg/800px-Da15_1_hic.jpg)
![](https://static.igem.org/mediawiki/2015/thumb/8/85/Da15_2_hic.jpg/800px-Da15_2_hic.jpg)
![](https://static.igem.org/mediawiki/2015/thumb/e/e2/Da15_3_hic.jpg/800px-Da15_3_hic.jpg)
![](https://static.igem.org/mediawiki/2015/8/80/Da15_hic.png)
![](https://static.igem.org/mediawiki/2015/c/c7/Da15_hic_heat_test.png)
[edit]
Categories
Parameters
//awards/part_collection/2015
None |