Team:Illinois/MicA
From 2009.igem.org
(→MicA Target-GFP Fusion) |
|||
Line 3: | Line 3: | ||
[https://2009.igem.org/Team:Illinois/sRNA_Library Back to sRNA Library Team page] | [https://2009.igem.org/Team:Illinois/sRNA_Library Back to sRNA Library Team page] | ||
+ | |||
+ | <html> | ||
+ | <div id="uicontentbox"> | ||
+ | </html> | ||
== '''MicA''' == | == '''MicA''' == |
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.