Team:Paris/Notebook/Oligo
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+ | {{Paris/Menu}} | ||
+ | |||
+ | ==Designed Oligonucleotides== | ||
{| | {| | ||
- | |- style="background: # | + | |- style="background: #649CD7;" |
- | ! colspan="5" style="background: # | + | ! colspan="5" style="background: #649CD7;" | Table of Oligos |
- | |- style="background: # | + | |- style="background: #649CD7; text-align: center;" |
|width=5%| Number | |width=5%| Number | ||
|width=10%| Name | |width=10%| Name | ||
|width=40%| Sequence | |width=40%| Sequence | ||
|width=5%| Length | |width=5%| Length | ||
- | |width= | + | |width=5%| Tm(°C) |
- | |- style="background: # | + | |width=35%| Comments |
- | | style="background: # | + | |- style="background: #dddddd;" |
+ | | style="background: #D4E2EF;" |O100 | ||
| FlgA-F | | FlgA-F | ||
+ | | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGAGCATATCTCCTCCGCAGGTATCAAAAT | ||
+ | | 58 | ||
| | | | ||
- | + | | | |
- | | | + | |- style="background: #dddddd;" |
- | |- style="background: # | + | | style="background: #D4E2EF;" | O101 |
- | | style="background: # | + | |
| FlgA-R | | FlgA-R | ||
- | | | + | | GTTTCTTCCTGCAGCGGCCGCTACTAGTAACAGTATCGCGATGATCGCCACGCTACGT |
- | | | + | | 58 |
| | | | ||
- | |- style="background: # | + | | |
- | | style="background: # | + | |- style="background: #dddddd;" |
+ | | style="background: #D4E2EF;" | O102 | ||
| FlgB-F | | FlgB-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGACAGTATCGCGATGATCGCCACGCTACG |
- | | | + | | 58 |
| | | | ||
- | |- style="background: # | + | | |
- | | style="background: # | + | |- style="background: #dddddd;" |
+ | | style="background: #D4E2EF;" | O103 | ||
| FlgB-R | | FlgB-R | ||
- | | | + | | GTTTCTTCCTGCAGCGGCCGCTACTAGTAAGCATATCTCCTCCGCAGGTATCAAAATT |
- | | | + | | 58 |
| | | | ||
- | |- style="background: # | + | | |
- | | style="background: # | + | |- style="background: #dddddd;" |
+ | | style="background: #D4E2EF;" | O104 | ||
| FlgK-F | | FlgK-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGCAGCAGATGAAATCGATAAGCGACAAGG |
- | | | + | | 58 |
| | | | ||
- | |- style="background: # | + | | |
- | | style="background: # | + | |- style="background: #dddddd;" |
+ | | style="background: #D4E2EF;" | O105 | ||
| FlgK-R | | FlgK-R | ||
- | | | + | | GTTTCTTCCTGCAGCGGCCGCTACTAGTAAACGCCGCCTGGGCCGCGTTCAGTCCGCT |
- | | | + | | 58 |
| | | | ||
- | |- style="background: # | + | | |
- | | style="background: # | + | |- style="background: #dddddd;" |
+ | | style="background: #D4E2EF;" | O106 | ||
| FlgM-F | | FlgM-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGAGCGGCGTTGTTGATGCAGATGGGAATA |
- | | | + | | 58 |
| | | | ||
- | |- style="background: # | + | | |
- | | style="background: # | + | |- style="background: #dddddd;" |
+ | | style="background: #D4E2EF;" | O107 | ||
| FlgM-R | | FlgM-R | ||
- | | | + | | GTTTCTTCCTGCAGCGGCCGCTACTAGTACGAAGTGCGATCAATACTCATGGTTTATT |
- | | | + | | 58 |
+ | | | ||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O108 |
| FlhB-F | | FlhB-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGCCACGTCATATCAGGCGGTCTGATAAGG |
- | | | + | | 58 |
+ | | | ||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O109 |
| FlhB-R | | FlhB-R | ||
- | | | + | | GTTTCTTCCTGCAGCGGCCGCTACTAGTAGTTTTGTCGTCGCTCTCGTCAGACACGTC |
- | | | + | | 58 |
+ | | | ||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O110 |
| FlhDC-Uri-F | | FlhDC-Uri-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGTTGTATGTGCGTGTAGTGACGAGTACAG |
- | | | + | | 58 |
| | | | ||
- | |- style="background: # | + | |Don't amplify the both OmpR binding site |
- | | style="background: # | + | |- style="background: #dddddd;" |
+ | | style="background: #D4E2EF;" | O111 | ||
| FlhDC-Total-F | | FlhDC-Total-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGTCATTTTTGCTTGCTAGCGTACGGAAAA |
- | | | + | | 58 |
| | | | ||
- | |- style="background: # | + | |Amplify the both OmpR binding site |
- | | style="background: # | + | |- style="background: #dddddd;" |
+ | | style="background: #D4E2EF;" | O112 | ||
| FlhDC(rbs)-R | | FlhDC(rbs)-R | ||
- | | | + | | GTTTCTTCCTGCAGCGGCCGCTACTAGTATCCCACCCAGAATAACCAACTTTAT |
- | | | + | | 54 |
| | | | ||
- | |- style="background: # | + | |Amplify the natural rbs of FlhD |
- | | style="background: # | + | |- style="background: #dddddd;" |
- | | FlhDC( | + | | style="background: #D4E2EF;" | O113 |
- | | | + | | FlhDC(rbs)-R |
- | | | + | | GTTTCTTCCTGCAGCGGCCGCTACTAGTACAGAATAACCAACTTTATTTTTATG |
+ | | 54 | ||
| | | | ||
- | |- style="background: # | + | |Amplify the natural rbs of FlhD |
- | | style="background: # | + | |- style="background: #dddddd;" |
- | | FliA-PCR1-F | + | | style="background: #D4E2EF;" | O114 |
- | | | + | | xx FliA-PCR1-F |
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGCCTGATTAACTGAGACTGACGGCAACGC |
+ | | 58 | ||
| | | | ||
- | |||
- | |||
- | |||
- | |||
- | |||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O115 |
- | | FliA- | + | | xx FliA-PCR1-R |
- | | | + | | CAGCCCTGCGTTATATGAGTTATCGGCATGATTATCCGTTTCCGCAGGGTTTTTAATCG |
- | | | + | | 59 |
| | | | ||
- | |||
- | |||
- | |||
- | |||
- | |||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O116 |
+ | | xx FliA-PCR2-F | ||
+ | | GAAACGGATAATCATGCCGATAACTCATATAACGCAGGGCTG | ||
+ | | 42 | ||
+ | | | ||
+ | | | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O117 | ||
+ | | xx FliA-PCR2-R | ||
+ | | GTTTCTTCCTGCAGCGGCCGCTACTAGTACCATTACACCTTCA | ||
+ | GCGGTATAGAGTGAACTCACGATAAA | ||
+ | | 69 | ||
+ | | | ||
+ | | | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O118 | ||
| FliC-F | | FliC-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGCCAGCGATGAAATACTTGCCATGCGATT |
- | | | + | | 58 |
| | | | ||
- | |- style="background: # | + | | |
- | | style="background: # | + | |- style="background: #dddddd;" |
+ | | style="background: #D4E2EF;" | O119 | ||
| FliC-R | | FliC-R | ||
- | | | + | | GTTTCTTCCTGCAGCGGCCGCTACTAGTACAGCGAGAGGCTGTTGGTATTAATGACTT |
- | | | + | | 58 |
| | | | ||
- | |- style="background: # | + | | |
- | | style="background: # | + | |- style="background: #dddddd;" |
+ | | style="background: #D4E2EF;" | O120 | ||
| FliE-F | | FliE-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGGTCGCATTCATCGCGCACCTCGTGGCTG |
- | | | + | | 58 |
| | | | ||
- | |- style="background: # | + | | |
- | | style="background: # | + | |- style="background: #dddddd;" |
+ | | style="background: #D4E2EF;" | O121 | ||
| FliE-R | | FliE-R | ||
- | | | + | | GTTTCTTCCTGCAGCGGCCGCTACTAGTACTGGCTGATAACCCCTTCAATCCCCTGTA |
- | | | + | | 58 |
+ | | | ||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O122 |
| FliF-F | | FliF-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGCTGGCTGATAACCCCTTCAATCCCCTGT |
- | | | + | | 58 |
+ | | | ||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O123 |
| FliF-R | | FliF-R | ||
- | | | + | | GTTTCTTCCTGCAGCGGCCGCTACTAGTAGTCGCATTCATCGCGCACCTCGTGGCTGA |
- | | | + | | 58 |
| | | | ||
- | |||
- | |||
- | |||
- | |||
- | |||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O124 |
- | | | + | | Oligo-pfliL-Forward-TRO |
- | | | + | | TCGAATTCGCGGCCGCTTCTAGAGCAAGGGCGTGTAACAGGCAAC |
- | | | + | | 45 |
+ | | 60°C | ||
+ | | | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O125 | ||
+ | | Oligo-pfliL-Reverse-TRO | ||
+ | | TCCTGCAGCGGCCGCTACTAGTAGTCATGTGTTGCGGTCTTCCTGTG | ||
+ | | 47 | ||
+ | | 62°C | ||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O126 |
| Gene-EnvZ-F | | Gene-EnvZ-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGATGAGGCGATTGCGCTTCTCGCCAC |
- | | | + | | 53 |
| | | | ||
- | |||
- | |||
- | |||
- | |||
- | |||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O127 |
+ | | Gene-EnvZ-R-TRO | ||
+ | | GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTACCCTTCTTTTGTCGTGCCCTGCGCC | ||
+ | | 60 | ||
+ | | 68°C | ||
+ | | | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O128 | ||
| Gene-FlgM-F | | Gene-FlgM-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGATGAGTATTGATCGCACTTCGCCTC |
- | | | + | | 53 |
| | | | ||
- | |||
- | |||
- | |||
- | |||
- | |||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O129 |
+ | | Gene-FlgM-R-TRO | ||
+ | | GTTTCTTCGAATTCGCGGCCGCTTCTAGTTATTAGTTACTCTGCAAGTCTTGCTGCGCT | ||
+ | | 59 | ||
+ | | 59°C | ||
+ | | | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O130 | ||
| Gene-FlhC-F | | Gene-FlhC-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGATGAGTGAAAAAAGCATTGTTCAGG |
- | | | + | | 53 |
| | | | ||
- | |||
- | |||
- | |||
- | |||
- | |||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O131 |
+ | | Gene-FlhC-R-TRO | ||
+ | | GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAAACAGCCTGTACTCTCTGTTCATCC | ||
+ | | 60 | ||
+ | | 60°C | ||
+ | | | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O132 | ||
| Gene-FlhD-F | | Gene-FlhD-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGATGCATACCTCCGAGTTGCTGAAAC |
- | | | + | | 53 |
| | | | ||
- | |- style="background: # | + | |Don't amplify the natural rbs of FlhD |
- | | style="background: # | + | |- style="background: #dddddd;" |
- | | Gene-FlhD-R | + | | style="background: #D4E2EF;" | O133 |
- | | | + | | Gene-FlhD-R-TRO |
- | | | + | | GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAGGCCCTTTTCTTGCGCAGCGCTTCT |
+ | | 60 | ||
+ | | 68°C | ||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O134 |
| Gene-FlhDCrbs-F | | Gene-FlhDCrbs-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGACCGCATAAAAATAAAGTTGGTTAT |
- | | | + | | 53 |
| | | | ||
- | |- style="background: # | + | |Amplify the natural rbs of FlhD so that we have FlhD and FlhC with both their natural rbs |
- | | style="background: # | + | |- style="background: #dddddd;" |
- | | Gene-FliA-F | + | | style="background: #D4E2EF;" | O135 |
- | | | + | | xx Gene-FliA-F |
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGATGAATTCACTCTATACCGCTGAAG |
+ | | 53 | ||
+ | | | ||
+ | |Don't Amplify the natural rbs of FliA | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O136 | ||
+ | | xx Gene-FliA-R | ||
+ | | GTTTCTTCGAATTCGCGGCCGCTTCTAGTTATTATAACTTACCCAGTTTAGTGCGTAAC | ||
+ | | 59 | ||
| | | | ||
- | |||
- | |||
- | |||
- | |||
- | |||
| | | | ||
- | |||
- | |||
- | |||
- | |||
- | |||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O137 |
+ | | xx Gene-FliArbs-F | ||
+ | | GTTTCTTCGAATTCGCGGCCGCTTCTAGTAATCATGCCGATAACTCATATAAC | ||
+ | | 53 | ||
+ | | | ||
+ | |Amplify the natural rbs of FliA | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O138 | ||
| Gene-OmpR-F | | Gene-OmpR-F | ||
- | | | + | | GTTTCTTCGAATTCGCGGCCGCTTCTAGATGCAAGAGAACTACAAGATTCTGG |
- | | | + | | 53 |
+ | | | ||
| | | | ||
- | |- style="background: # | + | |- style="background: #dddddd;" |
- | | style="background: # | + | | style="background: #D4E2EF;" | O139 |
| Gene-OmpR-R | | Gene-OmpR-R | ||
+ | | GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTATGCTTTAGAGCCGTCCGGTACAAAG | ||
+ | | 60 | ||
+ | | 62°C | ||
+ | | | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O140 | ||
+ | | PlasTest_GFP_Rv | ||
+ | | CCCTGCAGTTAATTAATATAAACGCAGAAAGGCCAC | ||
+ | | 36 | ||
+ | |54 | ||
+ | |Amplify E0240 with a PacI and a PstI restriction site at the end | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O141 | ||
+ | | PTst_GFPrbs+_F | ||
+ | | GCCTGCAGTCACACAGGAAAGTACTAGATGC | ||
+ | | 31 | ||
+ | |56 | ||
+ | |Amplify E0240 with the promoter and a PstI restriction site at the beginning | ||
+ | |- style="background: #dddddd;" | ||
+ | | style="background: #D4E2EF;" | O142 | ||
+ | | PTst_GFPrbs-_F | ||
+ | | GCCTGCAGATGCGTAAAGGAGAAGAACTTTTC | ||
+ | | 32 | ||
+ | | 55 | ||
+ | | 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> | ||
+ | <br> |
Latest revision as of 02:50, 27 September 2008
Designed Oligonucleotides
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