Team:Illinois/MicA

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Latest revision as of 02:43, 22 October 2009

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MicA

Primers Used: for the sRNA gene we used Forward Primer: GAA AGA CGC GCA TTT GTT ATC

Temp: (4*9) + (2*12) = 60 degrees

Second homology sequence: TTC CAG CCA CAC CGC AAA CGG ((TTCGGTATCA))

Reverse complement: CCG TTT GCG GTG TGG CTG G
Temp: (4*13) + (2*6) = 64 degrees
Cut site and overhang: GTTTTT TCTAGA

Reverse Primer: GTTTTT TCTAGA CCG TTT GCG GTG TGG CTG G

For the Target Sequence we used the primers from the Urban and Vogel paper: JV-0432 and JV-0433

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.

@@ Line 2: / Line 2: @@
 {{Illinois}}
-== '''MicA Target-GFP Fusion''' ==
+[https://2009.igem.org/Team:Illinois/sRNA_Library Back to sRNA Library Team page]
-'''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.
+<html>
+<div id="uicontentbox">
+</html>
-'''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"
+== '''MicA''' ==
-'''Recipe(s) Used:'''
+'''Primers Used: for the sRNA gene we used
+Forward Primer: GAA AGA CGC GCA TTT GTT ATC
+ Temp: (4*9) + (2*12) = 60 degrees
+Second homology sequence: TTC CAG CCA CAC CGC AAA CGG ((TTCGGTATCA))
+ Reverse complement: CCG TTT GCG GTG TGG CTG G
+ Temp: (4*13) + (2*6) = 64 degrees
+ Cut site and overhang: GTTTTT TCTAGA
+Reverse Primer: GTTTTT TCTAGA CCG TTT GCG GTG TGG CTG G
-'''Primers Used:'''
+For the Target Sequence we used the primers from the Urban and Vogel paper: JV-0432 and JV-0433
+'''
-== '''June 12''' ==
-Journal entry text goes here.
 == '''June 16'''==
@@ Line 20: / Line 26: @@
 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.
-[[Image:illinoisgelofmicA/OmpA.jpg]]
+[[Image: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.