Difference between revisions of "Team:Glasgow/Project/Overview/Terminator"
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| + | Protocols for CaCl<sub>2</sub> competent cells, transformation, miniprep, restriction digest, gel electrophoresis, ethidium bromide staining, Azure A staining, gel extraction, oligo annealing, and ligation available on our <a href="https://2015.igem.org/Team:Glasgow/Project/Overview/Protocols">Protocols</a> page. Fluorescence measurements taken as documented on our <a href="https://2015.igem.org/Team:Glasgow/Interlab">Interlab Study</a> page. | ||
<b> Figure 1. The terminator testing plasmid used was pSB1A10.</b> | <b> Figure 1. The terminator testing plasmid used was pSB1A10.</b> | ||
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Revision as of 23:08, 17 September 2015
Terminator
Home > Project > Terminator
Summary
Aim: To characterise the T1 terminator from E. coli rrnB, K1725081, for submission to the registry. Results Overview: K1725081 was successful in terminating transcription to the same level as B0010 and B0015. Basic Parts submitted: • BBa_K1725081
Introduction
In prokaryotes, such as E. coli, terminators occur at the end of a coding region or operon, and stop transcription. K1725081 was used in our Bistable Switch.
Methods
To characterise K1725081, the plasmid backbone pSB1A10 was used (figure 1). pSB1A10 contains I0500 (pBAD/araC), B0034 (strong ribosome binding site), E0040 (GFP), BioBrick prefix, BioBrick suffix, B0034, E1010 (RFP). With no terminator between the BioBrick prefix and suffix, cells transformed with the plasmid would fluoresce both green and red, however, when a terminator is inserted between the prefix and suffix, RFP expression is reduced, and the cells would fluoresce green.
Protocols for CaCl2 competent cells, transformation, miniprep, restriction digest, gel electrophoresis, ethidium bromide staining, Azure A staining, gel extraction, oligo annealing, and ligation available on our Protocols page. Fluorescence measurements taken as documented on our Interlab Study page.
Figure 1. The terminator testing plasmid used was pSB1A10.
Results
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