IGEM:IMPERIAL/2009/Primers: Difference between revisions
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Backward: <b>GGACTAGTA</b>TTAGTCTTTATCTGCCGG 51.4C(without) and 63.4(with)<br> | Backward: <b>GGACTAGTA</b>TTAGTCTTTATCTGCCGG 51.4C(without) and 63.4(with)<br> | ||
<br> | <br> | ||
Strains to use:DH10B | Strains to use:DH10B/TOP10 | ||
<br> | <br> |
Revision as of 07:45, 10 August 2009
Primers to be used for PCR
Considerations for primer design http://www.premierbiosoft.com/tech_notes/PCR_Primer_Design.html
Use this website to calculate temperatures and modify primers and this one for reverse complement.
RcsB Sequence:
ATGAACAATATGAACGTAATTATTGCCGATGACCATCCGATAGTCTTGTTCGGTATTCGCAAATCACTTG AGCAAATTGAGTGGGTGAATGTTGTCGGCGAATTTGAAGACTCTACAGCACTGATCAACAACCTGCCGAA ACTGGATGCGCATGTGTTGATTACCGATCTCTCCATGCCTGGCGATAAGTACGGCGATGGCATTACCTTA ATCAAGTACATCAAGCGCCATTTCCCAAGCCTGTCGATCATTGTTCTGACTATGAACAACAACCCGGCGA TTCTTAGTGCGGTATTGGATCTGGATATCGAAGGGATCGTGCTGAAACAAGGTGCACCGACCGATCTGCC GAAAGCTCTCGCCGCGCTGCAGAAAGGGAAGAAATTTACCCCGGAAAGCGTTTCTCGCCTGTTGGAAAAA ATCAGTGCTGGTGGTTACGGTGACAAGCGTCTCTCGCCAAAAGAGAGTGAAGTTCTGCGCCTGTTTGCGG AAGGCTTCCTGGTGACCGAGATCGCTAAAAAGCTGAACCGCAGTATTAAAACCATCAGTAGCCAGAAGAA ATCTGCGATGATGAAGCTGGGTGTCGAGAACGATATCGCCCTGCTGAATTATCTCTCTTCAGTGACCTTA AGTCCGGCAGATAAAGACTAA
Forward: GCTCTAGATGAACAATATGAACGTAATTATTG 53.1C(without) and 61.8(with)
Backward: GGACTAGTATTAGTCTTTATCTGCCGG 51.4C(without) and 63.4(with)
Strains to use:DH10B/TOP10
OtsB Sequence:
ATGACAGAACCGTTAACCGAAACCCCTGAACTATCCGCGAAATATGCCTGGTTTTTTGATCTTGATG GAACGCTGGCGGAAATCAAACCGCATCCCGATCAGGTCGTCGTGCCTGACAATATTCTGCAAGGACT ACAGCTACTGGCAACCGCAAGTGATGGTGCATTGGCATTGATATCAGGGCGCTCAATGGTGGAGCTT GACGCACTGGCAAAACCTTATCGCTTCCCGTTAGCGGGCGTGCATGGGGCGGAGCGCCGTGACATCA ATGGTAAAACACATATCGTTCATCTGCCGGATGCGATTGCGCGTGATATTAGCGTGCAACTGCATAC AGTCATCGCTCAGTATCCCGGCGCGGAGCTGGAGGCGAAAGGGATGGCTTTTGCGCTGCATTATCGT CAGGCTCCGCAGCATGAAGACGCATTAATGACATTAGCGCAACGTATTACTCAGATCTGGCCACAAA TGGCGTTACAGCAGGGAAAGTGTGTTGTCGAGATCAAACCGAGAGGTACCAGTAAAGGTGAGGCAAT TGCAGCTTTTATGCAGGAAGCTCCCTTTATCGGGCGAACGCCCGTATTTCTGGGCGATGATTTAACC GATGAATCTGGCTTCGCAGTCGTTAACCGACTGGGCGGAATGTCAGTAAAAATTGGCACAGGTGCAA CTCAGGCATCATGGCGACTGGCGGGTGTGCCGGATGTCTGGAGCTGGCTTGAAATGATAACCACCGC ATTACAACAAAAAAGAGAAAATAACAGGAGTGATGACTATGAGTCGTTTAGTCGTAGTATCTAA
Forward: GCTCTAGATGACAGAACCGTTAACCG 54.5C (without) & 64.8C (with)
Backward: GGACTAGTATTAGATACTACGACTAAACGAC 54.7C (without) & 64.2C (with)
B3023 Sequence:
ATGACAAACCTGACACTGGATGTAAACATTATCGATTTCCCATCAATACCTGTGGCGATGTTGCCG CACCGCTGTAGCCCTGAATTGCTCAACTACAGCGTGGCGAAATTTATCATGTGGCGTAAAGAGACG GGGCTTTCTCCTGTTAACCAAAGCCAGACTTTTGGCGTCGCCTGGGACGACCCTGCCACCACCGCA CCGGAAGCGTTTCGCTTTGATATCTGCGGCAGCGTTAGCGAACCGATTCCCGATAATCGTTATGGT GTGAGCAATGGTGAACTTACCGGTGGACGTTATGCCGTGGCCCGCCACGTTGGCGAGCTGGACGAT ATTTCACACACGGTATGGGGCATCATTCGCCACTGGCTGCCTGCAAGCGGCGAGAAAATGCGTAAA GCACCGATTCTGTTTCACTACACCAATCTTGCCGAAGGGGTGACAGAGCAGCGACTGGAAACGGAT GTTTATGTGCCGTTGGCGTGA
Forward: GCTCTAGATGACAAACCTGACACTG 51.4C(without) and 63.0(with)
Backward: GGACTAGTATCACGCCAACGGCAC 53.3C(without) and 66.1C(with)
WaaL Sequence:
ATGCTAACATCCTTTAAACTTCATTCATTGAAACCTTACACTCTGAAATCATCAATGATTTTAGAG ATAATAACTTATATATTATGTTTTTTTTCAATGATAATTGCATTCGTCGATAATACTTTCAGCATA AAAATATATAATATCACTGCTATAGTTTGCTTATTGTCACTAATTTTACGTGGCAGACAAGAAAATT ATAATATAAAAAACCTTATTCTTCCCCTTTCTATATTTTTAATAGGCTTGCTTGATTTAATTTGGTA TTCTGCGTTTAAAGTAGATAATTCGCCATTTCGTGCTACTTACCATAGTTATTTAAATACTGCCAAA ATATTTATATTTGGTTCTTTTATTGTTTTCTTGACACTAACTAGCCAGCTAAAATCAAAAAAAGAGA GTGTATTATACACTTTGTATTCTCTGTCATTTCTAATTGCTGGATATGCAATGTATATTAATAGCAT TCATGAAAATGACCGCATTTCTTTTGGTGTAGGAACGGCAACAGGAGCAGCATATTCAACAATGCTA ATAGGGATAGTTAGTGGCGTTGCGATTCTTTATACTAAGAAAAATCATCCTTTTTTATTTTTATTAA ATAGTTGCGCGGTACTTTATGTTCTGGCGCTAACACAAACCAGAGCAACCCTACTCCTGTTCCCTAT AATTTGTGTTGCTGCATTAATAGCTTATTATAATAAATCACCCAAGAAATTCACTTCCTCTATTGTT CTACTAATTGCTATATTAGCTAGCATTGTTATTATATTTAATAAACCAATACAGAATCGCTATAATG AAGCATTAAATGACTTAAACAGTTATACCAATGCTAATAGTGTTACTTCCCTAGGTGCAAGACTGGC AATGTACGAAATTGGTTTAAATATATTCATAAAGTCACCTTTTTCATTTAGATCAGCAGAGTCACGC GCTGAAAGTATGAATTTGTTAGTTGCAGAACACAATAGGCTAAGAGGGGCATTGGAGTTTTCTAACG TACATCTACATAATGAGATAATTGAAGCAGGGTCACTGAAAGGTCTGATGGGAATTTTTTCCACACT TTTCCTCTATTTTTCACTATTTTATATAGCATATAAAAAACGAGCTTTGGGTTTGTTGATATTAACG CTTGGCATTGTGGGGATTGGACTCAGTGATGTGATCATATGGGCACGCAGCATTCCAATTATCATTA TATCCGCTATAGTCCTCTTACTCGTCATTAATAATCGTAACAATACAATTAATTAA
Forward: GCTCTAGATGCTAACATCCTTTAAACTTC 52.8C(without) and 62.4(with)
Backward: GGACTAGTATTAATTAATTGTATTGTTACGATTATTAATG 54.9C(without) and 62.3(with)
OtsA Sequence:
ATGAGTCGTTTAGTCGTAGTATCTAACCGGATTGCACCACCAGACGAGCACGCCGCCAGTGCCGGTGGCCTTGC CGTTGGCATACTGGGGGCACTGAAAGCCGCAGGCGGACTGTGGTTTGGCTGGAGTGGTGAAACAGGGAATGAGG ATCAGCCGCTAAAAAAGGTGAAAAAAGGTAACATTACGTGGGCCTCTTTTAACCTCAGCGAACAGGACCTTGAC GAATACTACAACCAATTCTCCAATGCCGTTCTCTGGCCCGCTTTTCATTATCGGCTCGATCTGGTGCAATTTCA GCGTCCTGCCTGGGACGGCTATCTACGCGTAAATGCGTTGCTGGCAGATAAATTACTGCCGCTGTTGCAAGACG ATGACATTATCTGGATCCACGATTATCACCTGTTGCCATTTGCGCATGAATTACGCAAACGGGGAGTGAATAAT CGCATTGGTTTCTTTCTGCATATTCCTTTCCCGACACCGGAAATCTTCAACGCGCTGCCGACATATGACACCTT GCTTGAACAGCTTTGTGATTATGATTTGCTGGGTTTCCAGACAGAAAACGATCGTCTGGCGTTCCTGGATTGTC TTTCTAACCTGACCCGCGTCACGACACGTAGCGCAAAAAGCCATACAGCCTGGGGCAAAGCATTTCGAACAGAA GTCTACCCGATCGGCATTGAACCGAAAGAAATAGCCAAACAGGCTGCCGGGCCACTGCCGCCAAAACTGGCGCA ACTTAAAGCGGAACTGAAAAACGTACAAAATATCTTTTCTGTCGAACGGCTGGATTATTCCAAAGGTTTGCCAG AGCGTTTTCTCGCCTATGAAGCGTTGCTGGAAAAATATCCGCAGCATCATGGTAAAATTCGTTATACCCAGATT GCACCAACGTCGCGTGGTGATGTGCAAGCCTATCAGGATATTCGTCATCAGCTCGAAAATGAAGCTGGACGAAT TAATGGTAAATACGGGCAATTAGGCTGGACGCCGCTTTATTATTTGAATCAGCATTTTGACCGTAAATTACTGA TGAAAATATTCCGCTACTCTGACGTGGGCTTAGTGACGCCACTGCGTGACGGGATGAACCTGGTAGCAAAAGAG TATGTTGCTGCTCAGGACCCAGCCAATCCGGGCGTTCTTGTTCTTTCGCAATTTGCGGGAGCGGCAAACGAGTT AACGTCGGCGTTAATTGTTAACCCCTACGATCGTGACGAAGTTGCAGCTGCGCTGGATCGTGCATTGACTATGT CGCTGGCGGAACGTATTTCCCGTCATGCAGAAATGCTGGACGTTATCGTGAAAAACGATATTAACCACTGGCAG GAGTGCTTCATTAGCGACCTAAAGCAGATAGTTCCGCGAAGCGCGGAAAGCCAGCAGCGCGATAAAGTTGCTAC CTTTCCAAAGCTTGCGTAG
Forward: GCTCTAGATGAGTCGTTTAGTCGTAG 52.4C(without) and 63.2(with)
Backward: GGACTAGTACTACGCAAGCTTTGGAAAG 54.5C(without) and 65.1(with)
PAH Sequence:
ATGTCCACTGCGGTCCTGGAAAACCCAGGCTTGGGCAGGAAACTCTCTGACTTTGGACAGGAAACAAGCTATAT TGAAGACAACTGCAATCAAAATGGTGCCATATCACTGATCTTCTCACTCAAAGAAGAAGTTGGTGCATTGGCCA AAGTATTGCGCTTATTTGAGGAGAATGATGTAAACCTGACCCACATTGAATCTAGACCTTCTCGTTTAAAGAAA GATGAGTATGAATTTTTCACCCATTTGGATAAACGTAGCCTGCCTGCTCTGACAAACATCATCAAGATCTTGAG GCATGACATTGGTGCCACTGTCCATGAGCTTTCACGAGATAAGAAGAAAGACACAGTGCCCTGGTTCCCAAGAA CCATTCAAGAGCTGGACAGATTTGCCAATCAGATTCTCAGCTATGGAGCGGAACTGGATGCTGACCACCCTGGT TTTAAAGATCCTGTGTACCGTGCAAGACGGAAGCAGTTTGCTGACATTGCCTACAACTACCGCCATGGGCAGCC CATCCCTCGAGTGGAATACATGGAGGAAGAAAAGAAAACATGGGGCACAGTGTTCAAGACTCTGAAGTCCTTGT ATAAAACCCATGCTTGCTATGAGTACAATCACATTTTTCCACTTCTTGAAAAGTACTGTGGCTTCCATGAAGAT AACATTCCCCAGCTGGAAGACGTTTCTCAATTCCTGCAGACTTGCACTGGTTTCCGCCTCCGACCTGTGGCTGG CCTGCTTTCCTCTCGGGATTTCTTGGGTGGCCTGGCCTTCCGAGTCTTCCACTGCACACAGTACATCAGACATG GATCCAAGCCCATGTATACCCCCGAACCTGACATCTGCCATGAGCTGTTGGGACATGTGCCCTTGTTTTCAGAT CGCAGCTTTGCCCAGTTTTCCCAGGAAATTGGCCTTGCCTCTCTGGGTGCACCTGATGAATACATTGAAAAGCT CGCCACAATTTACTGGTTTACTGTGGAGTTTGGGCTCTGCAAACAAGGAGACTCCATAAAGGCATATGGTGCTG GGCTCCTGTCATCCTTTGGTGAATTACAGTACTGCTTATCAGAGAAGCCAAAGCTTCTCCCCCTGGAGCTGGAG AAGACAGCCATCCAAAATTACACTGTCACGGAGTTCCAGCCCCTGTATTACGTGGCAGAGAGTTTTAATGATGC CAAGGAGAAAGTAAGGAACTTTGCTGCCACAATACCTCGGCCCTTCTCAGTTCGCTACGACCCATACACCCAAA GGATTGAGGTCTTGGACAATACCCAGCAGCTTAAGATTTTGGCTGATTCCATTAACAGTGAAATTGGAATCCTT TGCAGTGCCCTCCAGAAAATAAAGTAA
Forward: GCTCTAGATGTCCACTGCGGTCC 54.3C (without) & 66.0C (with)
Backward: GGACTAGTATTACTTTATTTTCTGGAGGGC 54.0C (without) & 64.0C (with)
Cellulase Sequence:
ATGAAAAAAATTGTTTCTTTGGTTTGTGTGCTTGTGATGCTGGTAAGCATCTTAGGCTCGTTTTCAGTCGTA GCGGCATCACCGGTAAAAGGCTTTCAGGTATCGGGAACAAAGCTTTTGGATGCAAGCGGAAACGAGCTTGTA ATGAGGGGCATGCGTGATATTTCAGCAATAGATTTGGTTAAAGAAATAAAAATCGGATGGAATTTGGGAAAT ACTTTGGATGCTCCTACAGAGACTGCCTGGGGAAATCCAAGGACAACCAAGGCAATGATAGAAAAGGTAAGG GAAATGGGCTTTAATGCCGTCAGAGTGCCTGTTACCTGGGATACGCACATCGGACCTGCTCCGGACTATAAA ATTGACGAAGCATGGCTGAACAGAGTTGAGGAAGTGGTAAACTATGTTCTTGACTGCGGTATGTACGCGATC ATAAATGTTCACCATGACAATACATGGATTATACCTACATATGCCAATGAGCAAAGGAGTAAAGAAAAACTT GTAAAAGTTTGGGAACAAATAGCAACCCGTTTTAAAGATTATGACGACCATTTGTTGTTTGAGACAATGAAC GAACCGAGAGAAGTAGGTTCACCTATGGAATGGATGGGCGGAACGTATGAAAACCGAGATGTGATAAACAGA TTTAATTTGGCGGTTGTTAATACCATCAGAGCAAGCGGCGGAAATAACGATAAAAGATTCATACTGGTTCCG ACCAATGCGGCAACCGGCCTGGATGTTGCATTAAACGACCTTGTCATTCCGAACAATGACAGCAGAGTCATA GTATCCATACATGCTTATTCACCGTATTTCTTTGCTATGGATGTCAACGGAACTTCATATTGGGGAAGTGAC TATGACAAGGCTTCTCTTACAAGTGAACTTGATGCTATTTACAACAGATTTGTGAAAAACGGAAGGGCTGTA ATTATCGGAGAATTCGGAACCATTGACAAGAACAACCTGTCTTCAAGGGTGGCTCATGCCGAGCACTATGCA AGAGAAGCAGTTTCAAGAGGAATTGCTGTTTTCTGGTGGGATAACGGCTATTACAATCCGGGTGATGCAGAG ACTTATGCATTGCTGAACAGAAAAACTCTCTCATGGTATTATCCTGAAATTGTCCAGGCTCTTATGAGAGGT GCCGGCGTTGAACCTTTAGTTTCACCGACTCCTACACCTACATTAATGCCGACCCCCTCGCCCACGGTGACA GCAAATATTTTGTACGGTGACGTAAACGGGGACGGAAAAATAAATTCTACAGACTGTACAATGCTAAAGAGA TATATTTTGCGTGGCATAGAAGAATTCCCAAGTCCTAGCGGAATTATAGCCGCTGACGTAAATGCGGATCTG AAAATCAATTCCACCGACTTGGTATTGATGAAAAAATATCTACTGCGCTCAATAGACAAATTTCCTGCGGAG GATTCTCAAACACCTGATGAAGACAATCCGGGCATTTTGTATAACGGAAGATTCGATTTTTCAGATCCGAAC GGTCCGAAATGCGCCTGGTCCGGCAGCAATGTTGAGCTGAATTTTTACGGCACGGAAGCAAGTGTGACTATC AAATCCGGCGGTGAGAACTGGTTCCAGGCTATTGTAGACGGCAATCCTCTTCCTCCTTTTTCGGTTAACGCT ACTACCTCTACCGTAAAGCTTGTAAGCGGTCTTGCAGAAGGAGCTCATCATCTTGTATTGTGGAAGAGGACA GAGGCATCCTTGGGAGAAGTTCAGTTCCTTGGGTTTGATTTTGGTTCAGGAAAGCTTCTTGCCGCACCGAAG CCTTTGGAAAGAAAGATTGAGTTTATCGGAGACTCCATCACATGTGCATACGGAAATGAAGGAACAAGCAAG GAGCAGTCTTTTACACCGAAAAATGAAAACAGCTATATGTCTTATGCGGCAATTACAGCCCGTAATTTGAAT GCAAGTGCAAATATGATTGCGTGGTCCGGAATCGGACTTACCATGAACTACGGCGGAGCCCCCGGACCTCTT ATAATGGACCGTTATCCTTATACCCTTCCTTACAGCGGAGTCAGATGGGATTTTAGCAAATATGTGCCTCAG GTTGTTGTAATCAATCTTGGTACCAATGATTTTTCTACATCATTTGCAGATAAAACAAAGTTTGTAACGGCA TATAAAAACCTTATAAGTGAAGTTCGCAGGAACTATCCGGATGCCCATATATTCTGCTGTGTCGGTCCGATG CTTTGGGGAACGGGCCTGGATTTGTGCCGCAGTTATGTTACGGAAGTTGTAAATGATTGTAACAGAAGCGGG GATTTAAAGGTGTATTTTGTTGAGTTTCCGCAGCAGGACGGAAGCACCGGATACGGAGAAGACTGGCATCCA AGTATTGCCACCCACCAGCTGATGGCTGAGCGGCTTACTGCGGAAATAAAAAACAAGCTTGGATGGTAA
Forward: GCTCTAGATGAAAAAAATTGTTTCTTTGGTTTG 53.8C (without) & 62.0C (with)
Backward: GGACTAGTATTACCATCCAAGCTTGTTTTTTATTTC 57.4C (without) & 64.9C (with)
Dam Sequence:
ATGAAGAAAAATCGCGCTTTTTTGAAGTGGGCAGGGGGCAAGTATCCCCTGCTTGATGATATTAAA CGGCATTTGCCCAAGGGCGAATGTCTGGTTGAGCCTTTTGTAGGTGCCGGGTCGGTGTTTCTCAAC ACCGACTTTTCTCGTTATATCCTTGCCGATATCAATAGCGACCTGATCAGTCTCTATAACATTGTG AAGATGCGTACTGATGAGTACGTACAGGCCGCACGCGAGCTGTTTGTTCCCGAAACAAATTGCGCC GAGGTTTACTATCAGTTCCGCGAAGAGTTCAACAAAAGCCAGGATCCGTTCCGTCGGGCGGTACTG TTTTTATATTTGAACCGCTACGGTTACAACGGCCTGTGTCGTTACAATCTGCGCGGTGAGTTTAAC GTGCCGTTCGGCCGCTACAAAAAACCCTATTTCCCGGAAGCAGAGTTGTATCACTTCGCTGAAAAA GCGCAGAATGCCTTTTTCTATTGTGAGTCTTACGCCGATAGCATGGCGCGCGCAGATGATGCATCC GTCGTCTATTGCGATCCGCCTTATGCACCGCTGTCTGCGACCGCCAACTTTACGGCGTATCACACA AACAGTTTTACGCTTGAACAACAAGCGCATCTGGCGGAGATCGCCGAAGGTCTGGTTGAGCGCCAT ATTCCAGTGCTGATCTCCAATCACGATACGATGTTAACGCGTGAGTGGTATCAGCGCGCAAAATTG CATGTCGTCAAAGTTCGACGCAGTATAAGCAGCAACGGCGGCACACGTAAAAAGGTGGACGAACTG CTGGCTTTGTACAAACCAGGAGTCGTTTCACCCGCGAAAAAATAA
Forward: GCTCTAGATGAAGAAAAATCGCGCTTTTTTG 55.9C (without) & 64.2C (with)
Backward: GGACTAGTATTATTTTTTCGCGGGTGAAAC 54.0C (without) & 64.0C (with)