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Team:Illinois/MicA
From 2009.igem.org
Contents |
MicA Target-GFP Fusion
Purpose: to test the efficiency of sRNA repression. This will be accomplished by forming two plasmids by ligating the target sequence of the sRNA onto the pXG-10 plasmid behind the reporter gene GFP and ligating the sRNA gene into the pJU-334 DNA strand. These plasmids will then be transformed into E. coli and the fluorescence measured with a plate reader.
Protocol(s) Used: The protocol we used is that taken from Urban and Vogel, "A Green Fluorescent Protein (GFP)-Based Plasmid System to Study Post-Transcriptional Control of Gene Expression In Vivo"
Recipe(s) Used:
Primers Used:
June 12
Journal entry text goes here.
June 16
We used PCR to extract the sRNA gene: MicA and target sequence : ompA. from the e.coli chromosome. We then ran a gel to make sure that we had the right DNA fragments. Our results corresponded to our predictions.
June 17
We are completing a digestion of MicA, OmpA, and OmpF. Following the digestion we will incubate with SAP and then run a gel to verify the digestion. We will then extract the DNA for the sRNA target sequence and sRNA gene.
