Team:Illinois/MicA

From 2009.igem.org

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

Latest revision as of 02:43, 22 October 2009

Click to go to the Illinois home page



Back to sRNA Library Team page

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.

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.