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Team:SDU-Denmark/primers
From 2010.igem.org
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'''''Primers for FlhDC master operon''''' | '''''Primers for FlhDC master operon''''' | ||
| - | We needed to design 4 primers for this part. The FlhDC operon contains a PST1 restriction site, therefore we | + | We needed to design 4 primers for this part. The FlhDC operon contains a PST1 restriction site, therefore we wanted to introduce a silent mutation, C --> T, at position 822 in the operon. |
| - | ''Reverse primer w. mutation at position 13:'' 5’-CATAAGCTGCGGTCAAAGCTGCCAA -3’ (59) | + | ''Reverse primer w. mutation at position 13 (position 822):'' 5’-CATAAGCTGCGGTCAAAGCTGCCAA -3’ (59) |
| - | ''Forward primer w. mutation at position 13:'' 5’-TTGGCAGCTTTGACCGCAGCTTATG-3’ (59) | + | ''Forward primer w. mutation at position 13 (position 822):'' 5’-TTGGCAGCTTTGACCGCAGCTTATG-3’ (59) |
Revision as of 14:40, 8 July 2010
Primers
On this page we list all the primers we designed and used during the summer. Numbers in paranthesis is basic temperature (celcius)
Primers for FlhDC master operon
We needed to design 4 primers for this part. The FlhDC operon contains a PST1 restriction site, therefore we wanted to introduce a silent mutation, C --> T, at position 822 in the operon.
Reverse primer w. mutation at position 13 (position 822): 5’-CATAAGCTGCGGTCAAAGCTGCCAA -3’ (59)
Forward primer w. mutation at position 13 (position 822): 5’-TTGGCAGCTTTGACCGCAGCTTATG-3’ (59)
Lastly we added primers with prefix and suffix to the PCR result of the above:
Prefixprimer: 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGCATACCTCCGAGTTGCT-3' (Coding sequence (CS): 52) (Primer total (PT): 72)
Suffixprimer: 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTAAACAGCCTGTACTCTCT-3' (CS: 45) (PT: 72)
SopIItrs
Forward primer: 5’-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGGTGGGACTTACGACC-3’ (CS: 50) (PT: 73)
Reverse primer: 5’-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTAAAATGTTTCCCAGTTCT-3’ (CS: 70) (PT: 44)
