Difference between revisions of "Part:BBa K1111005:Experience"
GigiGoesGugu (Talk | contribs) (→Sequencing) |
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We sequenced both the Promoter and the superfolded GFP. Both sequencing results showed that there were no mutations in the sequences. | We sequenced both the Promoter and the superfolded GFP. Both sequencing results showed that there were no mutations in the sequences. | ||
− | [[Image: Team-EPF-Lausanne 1.3 Sequencing_Promoter.jpg|thumb|600px| | + | [[Image: Team-EPF-Lausanne 1.3 Sequencing_Promoter.jpg|thumb|600px|center|Figure 1: Sequencing result of the constitutive promoter using one foreward Primer that binds directly to the beginning of the promoter ]] |
− | [[Image: Team-EPF-Lausanne 1.3 Sequencing_GFP.jpg|thumb|600px| | + | [[Image: Team-EPF-Lausanne 1.3 Sequencing_GFP.jpg|thumb|600px|center|Figure 1: Sequencing result of the superfolded GFP using one foreward Primer that binds to the VF2 bindning site in the pSB1C3 plasmid]] |
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+ | ===OD Measurements=== | ||
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+ | We transformed cell with our plasmid and then measured their OD in different media with different pHs. We compared this data with the Fluorescence measurements and were able to conclude that the reason why GFP expression decreases at low pH is that the cells are actually dying. | ||
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+ | [[Image: Team-EPF-Lausanne 1.3_OD_Measurements.jpg|thumb|600px|center|Figure 1: OD measurements of transformed DH5 alpha cells in media at varying pH values.]] | ||
===User Reviews=== | ===User Reviews=== |
Revision as of 18:38, 2 October 2013
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_K1111005
We constructed this plasmid in order to use it as a positive control for promoters that are inducible and inserten in front of the same reporter gene (superfolded GFP from iGEM). As it is constitutively expressed, the fluorescence can be used at least as a qualitative comparison to promoters that are induced.
Gibson Assembly and Primers
We used Gibson Assembly to insert this constitutive promoter into the pSB1C3 Backbone. We used the biobrick BBa_I746908 where we remove the Arac promoter but left the superfolded GFP. This way, the latter was under control of our Cad promoter.
Sequencing
We sequenced both the Promoter and the superfolded GFP. Both sequencing results showed that there were no mutations in the sequences.
OD Measurements
We transformed cell with our plasmid and then measured their OD in different media with different pHs. We compared this data with the Fluorescence measurements and were able to conclude that the reason why GFP expression decreases at low pH is that the cells are actually dying.
User Reviews
UNIQ928d7067be4ee968-partinfo-00000000-QINU UNIQ928d7067be4ee968-partinfo-00000001-QINU