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Team:Illinois/MicA
From 2009.igem.org
(New page: {{UIUC09Temp}} {{Illinois}} == '''MicA Target-GFP Fusion''' == '''Purpose:''' '''Protocol(s) Used:''' (links to protocols page) '''Recipe(s) Used:''' '''Primers Used:''' == '''June...) |
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| - | + | [https://2009.igem.org/Team:Illinois/sRNA_Library Back to sRNA Library Team page] | |
| - | + | <html> | |
| + | <div id="uicontentbox"> | ||
| + | </html> | ||
| - | ''' | + | == '''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 | + | == '''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. | |
| + | |||
| + | [[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. | ||
Latest revision as of 02:43, 22 October 2009
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.
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.
