Team:Illinois/MicA

From 2009.igem.org

(Difference between revisions)
(MicA Target-GFP Fusion)
(June 16)
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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.   
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.   
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[[Image:illinoisgelofmicA/OmpA.jpg]]
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[[Image:UI09Gel2.jpg]]
=='''June 17'''==
=='''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.
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.

Revision as of 16:50, 17 June 2009

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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.

UI09Gel2.jpg

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.