Team:SDU-Denmark/primers
From 2010.igem.org
(Difference between revisions)
(→Primers) |
|||
Line 6: | Line 6: | ||
== '''Primers''' == | == '''Primers''' == | ||
- | + | <br> | |
- | + | On this page we list all the primers we designed and used during the summer. Numbers in paranthesis is basic temperature (celcius) <br><br> | |
- | 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''''' | '''''Primers for FlhDC master operon''''' | ||
- | + | <br> | |
- | 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. | + | 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.<br><br> |
- | + | ''Reverse primer w. mutation at position 13 (gene position 822):'' 5’-CATAAGCTGCGGTCAAAGCTGCCAA -3’ (59)<br><br> | |
- | + | ''Forward primer w. mutation at position 13 (gene position 822):'' 5’-TTGGCAGCTTTGACCGCAGCTTATG-3’ (59)<br><br> | |
- | ''Reverse primer w. mutation at position 13 (gene position 822):'' 5’-CATAAGCTGCGGTCAAAGCTGCCAA -3’ (59) | + | Lastly we added primers with prefix and suffix to the PCR result of the above:<br><br> |
- | + | [[FlhDC forward:]]<br> | |
- | ''Forward primer w. mutation at position 13 (gene position 822):'' 5’-TTGGCAGCTTTGACCGCAGCTTATG-3’ (59) | + | ''Prefixprimer:'' 5'-G TTT CTT CGA ATT CGC GGC CGC TTC TAG ATG CAT ACC TCC GAG TTG CTG AAA-3' (Coding sequence (CS): 56) (Primer total (PT): 72)<br><br> |
- | + | [[FlhDC reverse:]]<br> | |
- | + | ''Suffixprimer:'' 5'-GT TTC TTC CTG CAG CGG CCG CTA CTA TTA AAC AGC CTG TAC TCT CTG TTC-3' (CS: 56) (PT: 72)<br><br> | |
- | Lastly we added primers with prefix and suffix to the PCR result of the above: | + | |
- | + | ||
- | + | ||
- | [[FlhDC forward:]] | + | |
- | + | ||
- | ''Prefixprimer:'' 5'-G TTT CTT CGA ATT CGC GGC CGC TTC TAG ATG CAT ACC TCC GAG TTG CTG AAA-3' (Coding sequence (CS): 56) (Primer total (PT): 72) | + | |
- | + | ||
- | [[FlhDC reverse:]] | + | |
- | + | ||
- | ''Suffixprimer:'' 5'-GT TTC TTC CTG CAG CGG CCG CTA CTA TTA AAC AGC CTG TAC TCT CTG TTC-3' (CS: 56) (PT: 72) | + | |
- | + | ||
- | + | ||
'''''SopIItrs''''' | '''''SopIItrs''''' | ||
- | + | <br> | |
- | ''Forward primer:'' 5’-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGGTGGGACTTACGACC-3’ (CS: 50) (PT: 73) | + | ''Forward primer:'' 5’-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGGTGGGACTTACGACC-3’ (CS: 50) (PT: 73)<br><br> |
- | + | ''Reverse primer:'' 5’-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTAAAATGTTTCCCAGTTCT-3’ (CS: 70) (PT: 44) <br><br> | |
- | ''Reverse primer:'' 5’-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTAAAATGTTTCCCAGTTCT-3’ (CS: 70) (PT: 44) | + | |
</div> | </div> |
Revision as of 15:45, 12 July 2010