Team:Illinois/MicF

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Latest revision as of 02:43, 22 October 2009

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MicF

Primers Used: (5' -> 3')

Forward MicF (sRNA gene) primer: 5'p+GCTATCATCATTAACTTTATTTATTACCG
Reverse MicF (sRNA gene) primer: GTTTTTTCTAGAGGTAGCACAGAATAATGAAA
Forward OmpF (sRNA gene) primer: GTTTTTATGCATAGACACATAAAGACACCAAACTC
Reverse OmpF (sRNA gene) primer: GTTTTTGCTAGCAGGGACGATCACTG

MicF gene sequence:

GCT ATC ATC ATT AAC TTT ATT TAT TAC CGT CAT TCA TTT CTG AAT GTC TGT TTA CCC CTA TTT CAA CCG GAT GCC TCG CAT TCG GTT TTT TTT ACC CTT CTT TAC ACA CTT TTC ATT ATT CTG

MicF binding domain on OmpF:

AGA CAC ATA AAG ACA CCA AAC TCT CAT CAA TAG TTC CGT AAA TTT TTA TTG ACA GAA CTT ATT GAC GGC AGT GGC AGG TGT CAT AAA AAA AAC CAT GAG GGT AAT AAA TAA TGA TGA AGC GCA ATA TTC TGG CAG TGA TCG TCC CTG C

(Bold is where the primers anneal)

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 MicF and OmpF. Following the digestion we will incubate with Shrimp Alkanline Phosphatase 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}}
-== '''MicF Target-GFP Fusion''' ==
+[https://2009.igem.org/Team:Illinois/sRNA_Library Back to sRNA Library Team page]
-'''Purpose:''' MicF will be fused to the target sequence from the OmpF gene to the GFP gene on a low-copy plasmid and transform this into E. coli cells along with a high-copy plasmid carrying the MicF gene.  We expect to see translational repression of GFP by the small RNA MicF.
+<html>
+<div id="uicontentbox">
+</html>
-'''Protocol(s) Used:''' (links to protocols page)
+== '''MicF''' ==
-'''Recipe(s) Used:'''
 '''Primers Used:''' (5' -> 3')
@@ Line 18: / Line 18: @@
 MicF gene sequence:
-*GCT ATC ATC ATT AAC TTT ATT TAT TAC CGT CAT TCA TTT CTG AAT GTC TGT TTA CCC CTA TTT CAA CCG GAT GCC TCG CAT TCG GTT TTT TTT ACC CTT CTT TAC ACA CTT TTC ATT ATT CTG
+*'''GCT ATC ATC ATT AAC TTT ATT TAT TAC CG'''T CAT TCA TTT CTG AAT GTC TGT TTA CCC CTA TTT CAA CCG GAT GCC TCG CAT TCG GTT TTT TTT ACC CTT CTT TAC ACA CT'''T TTC ATT ATT CTG'''
 MicF binding domain on OmpF:
-*
+*'''AGA CAC ATA AAG ACA CCA AAC TC'''T CAT CAA TAG TTC CGT AAA TTT TTA TTG ACA GAA CTT ATT GAC GGC AGT GGC AGG TGT CAT AAA AAA AAC CAT GAG GGT AAT AAA TAA TGA TGA AGC GCA ATA TTC TGG '''CAG TGA TCG TCC CTG C'''
+(Bold is where the primers anneal)
 == '''June 16'''==