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From 2010.igem.org

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 (gene position 822): 5’-CATAAGCTGCGGTCAAAGCTGCCAA -3’ (59)

Forward primer w. mutation at position 13 (gene position 822): 5’-TTGGCAGCTTTGACCGCAGCTTATG-3’ (59)

Lastly we added primers with prefix and suffix to the PCR result of the above:

FlhDC forward:

Prefixprimer: 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGCATACCTCCGAGTTGCT-3' (Coding sequence (CS): 52) (Primer total (PT): 72)

FlhDC reverse:

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)