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Team:Waterloo/Notebook/Oligos
From 2009.igem.org
(Difference between revisions)
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|amplifies second half of alpha gal gene, adds KpnI site (fwd) || H2-KpnI-attP-F || CATGGTACCCCCCCAACTGAGAGAACTCAAAGGTTACCCCAGTTGGGGGCGTTTTACCTGGAGCTGG || | |amplifies second half of alpha gal gene, adds KpnI site (fwd) || H2-KpnI-attP-F || CATGGTACCCCCCCAACTGAGAGAACTCAAAGGTTACCCCAGTTGGGGGCGTTTTACCTGGAGCTGG || | ||
|- | |- | ||
| - | | | + | |amplifies second half of alpha gal gene, adds XbaI site (rev) || H2-XbaI-R || ATGTCTAGAGGTCAGCAGCGTCTGT || |
|- | |- | ||
| - | | | + | |Amplifes any biobrick, adds EcoRV end for ligation into EcoRV blunt ends (fwd) || Prefix-EcoRV-F || atcgaattcgcggccgcttctag || |
| + | |- | ||
| + | |Amplifes any biobrick, adds EcoRV end for ligation into EcoRV blunt ends (rev) || Suffix-EcoRV-R || atcctgcagcggccgctactagta || | ||
|- | |- | ||
|B0034 RBS primer, use with VF2 (fwd) || Prefix-RBS || GGCCGCTTCTAGAGAAAGAGG || 21 | |B0034 RBS primer, use with VF2 (fwd) || Prefix-RBS || GGCCGCTTCTAGAGAAAGAGG || 21 | ||
|- | |- | ||
|B0034 RBS primer, use with VR (rev)|| Suffix-RBS || TTTCTCCTCTTTCTCTAGATGCGG || 24 | |B0034 RBS primer, use with VR (rev)|| Suffix-RBS || TTTCTCCTCTTTCTCTAGATGCGG || 24 | ||
| + | |- | ||
| + | |ordered to add Plac and RBS to the cI ORF (project 2008) || Plac-RBS-F || GAATTCGCGGCCGCTTCTAGAGAATTGTGAGCGGATAACAATTGACATTGTGAGCGGATAA CAAGATACTGAGCACATACTAGAGAAAGAGGAGAAATACTAG || 103 | ||
| + | |- | ||
| + | |||
| + | |BBa sequencing/Colony PCR primer (fwd) || VF2 || TGCCACCTGACGTCTAAGAA || 20 | ||
| + | |- | ||
| + | |BBa sequencing/Colony PCR primer (rev) || VR || ATTACCGCCTTTGAGTGAGC || 20 | ||
|- | |- | ||
|T7 gene 5 primer, 3'-5' exonuclease, adds BBa prefix (fwd) || T7gene5-F || CAATGGAATTCGCGGCCGCTTCTAGATGATCGTTTCTGACATCGAA || 46 | |T7 gene 5 primer, 3'-5' exonuclease, adds BBa prefix (fwd) || T7gene5-F || CAATGGAATTCGCGGCCGCTTCTAGATGATCGTTTCTGACATCGAA || 46 | ||
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|RFP ORF (fwd; prefix-RBS for joining to transcriptional terminator by crossover PCR ) || RFP-RXO || GATGCCTGGCTCTAGTATTATTAAGCACCGGTGG || 34 | |RFP ORF (fwd; prefix-RBS for joining to transcriptional terminator by crossover PCR ) || RFP-RXO || GATGCCTGGCTCTAGTATTATTAAGCACCGGTGG || 34 | ||
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Revision as of 06:03, 20 October 2009
| Description | Oligo Name | Sequence | Length |
|---|---|---|---|
| amplifies first half of E. coli alpha gal gene adds SalI REN site (fwd) | HI-SalI-F | CATGTCGACATGATGTCTGCACCCAAAATT | |
| amplifies first half of alpha gal gene adds BamHI REN site (rev) | HI-BamHI-attP-R | ATGGGATCCCCCCCAACTGAGAGAACTCAAAGGTTACCCCAGTTGGGGGTAACGCAGCGTAGCTGG | |
| amplifies second half of alpha gal gene, adds KpnI site (fwd) | H2-KpnI-attP-F | CATGGTACCCCCCCAACTGAGAGAACTCAAAGGTTACCCCAGTTGGGGGCGTTTTACCTGGAGCTGG | |
| amplifies second half of alpha gal gene, adds XbaI site (rev) | H2-XbaI-R | ATGTCTAGAGGTCAGCAGCGTCTGT | |
| Amplifes any biobrick, adds EcoRV end for ligation into EcoRV blunt ends (fwd) | Prefix-EcoRV-F | atcgaattcgcggccgcttctag | |
| Amplifes any biobrick, adds EcoRV end for ligation into EcoRV blunt ends (rev) | Suffix-EcoRV-R | atcctgcagcggccgctactagta | |
| B0034 RBS primer, use with VF2 (fwd) | Prefix-RBS | GGCCGCTTCTAGAGAAAGAGG | 21 |
| B0034 RBS primer, use with VR (rev) | Suffix-RBS | TTTCTCCTCTTTCTCTAGATGCGG | 24 |
| ordered to add Plac and RBS to the cI ORF (project 2008) | Plac-RBS-F | GAATTCGCGGCCGCTTCTAGAGAATTGTGAGCGGATAACAATTGACATTGTGAGCGGATAA CAAGATACTGAGCACATACTAGAGAAAGAGGAGAAATACTAG | 103 |
| BBa sequencing/Colony PCR primer (fwd) | VF2 | TGCCACCTGACGTCTAAGAA | 20 |
| BBa sequencing/Colony PCR primer (rev) | VR | ATTACCGCCTTTGAGTGAGC | 20 |
| T7 gene 5 primer, 3'-5' exonuclease, adds BBa prefix (fwd) | T7gene5-F | CAATGGAATTCGCGGCCGCTTCTAGATGATCGTTTCTGACATCGAA | 46 |
| T7 gene 5 primer, 3'-5' exonuclease, adds BBa suffix (rev) | T7gene5-R | CAATTTCTGCAGCGGCCGCTACTAGTATCAGTGGCAAATCGCCCAATTAGG | 51 |
| Gene 6 ORF from T7 phage (fwd; Prefix adapter) | T7gene6-F | CAATGGAATTCGCGGCCGCTTCTAGATGGCACTTCTTGACCTTAAA | 46 |
| Gene 6 ORF from T7 phage (rev; Suffix adapter) | T7gene6-R | CAATTTCTGCAGCGGCCGCTACTAGTACTACGGTCTCCACAGG | 43 |
| PmeIR ORF (fwd) | PmeIR-F | ATCCGGAATTCGCGGCGCTTTCTAGATGACAACAAACTCCCCCTCAGACG | 50 |
| PmeIR ORF (rev) | PmelR-R | CGCAAAACTGCAGCGGCCGCTACTAGTATCAAACCGTACTTCGCCCCCGTTG | 52 |
| pRecA oligo (sense strand; produces BioBrick-compatible EcoRI/SpeI overhangs when annealed to recAR) | recAF | AATTCGCGGCCGCTTCTAGAGTACAAAACACTTGATACTGTATATATATACAGTATAATTGCTTCAACATA | 71 |
| pRecA oligo (antisense strand; produces BioBrick-compatible EcoRI/SpeI overhangs when annealed to recAR) | recAR | CTAGTATGTTGAAGCAATTATACTGTATATATATACAGTATCAAGTGTTTTGTACTCTAGAAGCGGCCGCG | 71 |
| {B0034}-{T7gene6 ORF} (fwd; for adding {Prefix-RBS} onto the gene 6 ORF by PCR) | RBST7g6-F | GAATTCCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGGCACTTCTTGACCTTAAAC | 61 |
| T7gene6 ORF (rev) | T7gs-R | CTGCAGCGGCCGCTACTAGTATTATTACGGTCTCCACAGGTAAATCTC | 48 |
| T7gene6 ORF rev; for joining to PmeI with crossover PCR | T7g6-RXO-RBS | CTAGTATTTCTCCTCTTTCTCTAGTATTATTACGGTCTCCACAGGTAAATCTC | 53 |
| PmeI ORF (fwd; for adding prefix-RBS) | RBSpmeI-F | GAATTCCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGACAACAAACTCCCCCTCAG | 61 |
| PmeI ORF (fwd; for adding prefix-RBS and joining to T7gene6 by crossover PCR) | RBSpmeI-FXO | TACTAGAGAAAGAGGAGAAATACTAGATGACAACAAACTCCCCCTCAG | 48 |
| PmeI ORF (rev; TAATAA stop codon-suffix) | pmeI-R | CTGCAGCGGCCGCTACTAGTATTATTAAACCGTACTTCGCCCCCG | 45 |
| Primer for part R0011 (Plac-PL hybrid promoter; fwd) | R0011-F | GAATTCCGGCCGCTTCTAGAGAATTGTGAGCGGATAAC | 38 |
| Primer for part R0011 (Plac-PL hybrid promoter; rev for joining to ORFs with RBS by crossover PCR) | R0011RBS-RXO | CTAGTATTTCTCCTCTTTCTCTAGTATGTGCTCAGTATCTTGTTATC | 47 |
| CI ORF (fwd; prefix-RBS) | RBSC0051-F | GAATTCCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGAGCACAAAAAAGAAACC | 59 |
| CI ORF (fwd; prefix-RBS for joining to R0011 promoter by crossover PCR) | RBSC0051-FXO | TACTAGAGAAAGAGGAGAAATACTAGATGAGCACAAAAAAGAAACC | 46 |
| CI ORF (rev; suffix adapter) | C0051-R | CTGCAGCGGCCGCTACTAGTATTATTAAGCTACTAAAGCG | 40 |
| CI ORF (rev; for joining to transcriptional terminator (b0015) by crossover PCR) | C0051-RXO | GATGCCTGGCTCTAGTATTATTAAGCTACTAAAGCG | 36 |
| Transcriptional terminator (B0015; fwd; for joining to ORFs by crossover PCR) | B0015-FXO | TAATAATACTAGAGCCAGGCATCAAATAAAACG | 33 |
| Transcriptional terminator (B0015; rev) | B0015-R | CTGCAGCGGCCGCTACTAGTATATAAACGCAGAAAGGCC | 39 |
| RFP ORF (fwd; prefix-RBS) | RFP-RBS-F | GAATTCCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGGCTTCCTCCGAAGACG | 58 |
| RFP ORF (fwd; prefix-RBS for joining to transcriptional terminator by crossover PCR ) | RFP-RXO | GATGCCTGGCTCTAGTATTATTAAGCACCGGTGG | 34 |
