Team:Paris/Notebook/Oligo
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==Designed Oligonucleotides== | ==Designed Oligonucleotides== | ||
{| | {| | ||
Line 107: | Line 105: | ||
|- style="background: #dddddd;" | |- style="background: #dddddd;" | ||
| style="background: #D4E2EF;" | O113 | | style="background: #D4E2EF;" | O113 | ||
- | | FlhDC( | + | | FlhDC(rbs)-R |
| GTTTCTTCCTGCAGCGGCCGCTACTAGTACAGAATAACCAACTTTATTTTTATG | | GTTTCTTCCTGCAGCGGCCGCTACTAGTACAGAATAACCAACTTTATTTTTATG | ||
| 54 | | 54 | ||
| | | | ||
- | | | + | |Amplify the natural rbs of FlhD |
|- style="background: #dddddd;" | |- style="background: #dddddd;" | ||
| style="background: #D4E2EF;" | O114 | | style="background: #D4E2EF;" | O114 | ||
Line 315: | Line 313: | ||
| GCCTGCAGATGCGTAAAGGAGAAGAACTTTTC | | GCCTGCAGATGCGTAAAGGAGAAGAACTTTTC | ||
| 32 | | 32 | ||
- | |55 | + | | 55 |
- | |Amplify E0240 without the promoter and a PstI restriction site at the beginning | + | | Amplify E0240 without the promoter and a PstI restriction site at the beginning |
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O143 | ||
+ | | G-PCRmut-1F | ||
+ | | GGCTGTTTATCGTGAACTCACTCTATACCG | ||
+ | | 30 | ||
+ | | 62 | ||
+ | | Mutate the EcoRI site of FliA ( without biobrick prefix) | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O144 | ||
+ | | P-FliArbs-R | ||
+ | | CTTCCTGCAGCGGCCGCTACTAGTATTAAACAGCCCTGCGTTATATG | ||
+ | | 47 | ||
+ | | 55 - 72 | ||
+ | | Amplify the rbs of FliA | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O145 | ||
+ | | P-xb-FliA-F | ||
+ | | GCTTCTAGAGCCTGATTAACTGAGACTGAC | ||
+ | | 30 | ||
+ | | 51 - 62 | ||
+ | | No EcoRI site on prefix for amplifying promoter | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O146 | ||
+ | | P-FliA-R | ||
+ | | GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATATGAGTTATCGGCATGAT | ||
+ | | 51 | ||
+ | | 51 - 71 | ||
+ | | | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O147 | ||
+ | | G-xb-FliA-F | ||
+ | | CTTCTAGAACGCAGGGCTGTTTATCGTG | ||
+ | | 28 | ||
+ | | 57 - 63 | ||
+ | | Amplify the gene with its natural rbs | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O148 | ||
+ | | G-mecoFliA-F | ||
+ | | TCGAATTCGCGGCCGCTTCTAGACAGGGCTGTTTATCGTGAACTCACTCTATA | ||
+ | | 53 | ||
+ | | 61 - 74 | ||
+ | | Oligo for second run PCR | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O149 | ||
+ | | G-spe-FliA-R | ||
+ | | GCTACTAGTATTATTATAACTTACCCAGTTTAGTGCG | ||
+ | | 37 | ||
+ | | 53 - 60 | ||
+ | | Oligo FliA for non-mutated sequence | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O150 | ||
+ | | G-FliA-R | ||
+ | | TCCTGCAGCGGCCGCTACTAGTATTATTATAACTTACCCAGTTTAGTGCG | ||
+ | | 50 | ||
+ | | 53 - 71 | ||
+ | | Oligo for complete mutated sequence - second run of PCR | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O151 | ||
+ | | PG-xb-FliA-R | ||
+ | | GCTTCTAGAATTATTATAACTTACCCAGTTTAGTGCG | ||
+ | | 37 | ||
+ | | 53 - 61 | ||
+ | | Oligo reverse with xbaI site for non mutated sequence | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O152 | ||
+ | | G-PCRmut-2R | ||
+ | | GGGCAGTCGAACTTGCAGGCGCAATGCTTC | ||
+ | | 30 | ||
+ | | 71 | ||
+ | | Oligo for first run of PCR, mutate PstI. | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O153 | ||
+ | | G-PCRmut-3F | ||
+ | | GAAGCATTGCGCCTGCAAGTTCGACTGCCC | ||
+ | | 30 | ||
+ | | 71 | ||
+ | | Oligo for fisrt run of PCR, mutate PstI. | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O154 | ||
+ | | pFlhDC-tto-F | ||
+ | | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGGTCGCTGTCGTTCTCAAAATCGGTG | ||
+ | | 55 | ||
+ | | 62 - 75 | ||
+ | | | ||
|} | |} | ||
|}<br style="clear:both" /> | |}<br style="clear:both" /> | ||
<br> | <br> | ||
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Latest revision as of 02:50, 27 September 2008
Designed Oligonucleotides
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