Team:ULB-Brussels/mat

Modelling : Introduction

Primers The following primers were produced by Sigma-Aldrich (option desalt) Yeast cloning: PKD46-FOR: 5’-  gagcaaaaggccagcaaaaggccaggaaccgtaaaaaggctgccacctgcatcgatttat-3’ PKD46-REV: 5’-cgtgagttttcgttccactgagcgtcagaccccgtagaaagagttttcgttccactgagc-3’ PFL-FOR: 5’-Gcctttttacggttcctggc-3’ PFL-REV: 5’-tttctacgggggtctgacgc-3’ PCP-FOR: 5’-tggctcttgtatctatcagtgaagcatcaagactaacaaatcagccaaacgtctcttcag-3’ PCP-REV: 5’-ggggctgtatgcacaaagcatcttctgttgagttaagaacttatatgcgtctatttatgtagg-3’ In green is the 40 homologous nucleotides need for the yeast cloning. Yeast cloning verification: FLP/CI-FOR: 5’-Acatggcgagttttgacgag-3’ FLP/CI-REV: 5’-accacactagagaacatactg-3’ pINDEL sequencing: pID-seq1: 5’-aggatcttcacctagatcctt-3’ pID-seq2:  5’-gatgggctagtcaatgataatta-3’ pID-seq3: 5’-ccgttacgtaggtaggaatc-3’ pID-seq4: 5’-agatggggatggggcagtc-3’ pID-seq5: 5’-gatttcggatcaacgttcttaat-3’ pID-seq6: 5’-caatcactttcgtctactcc-3’ pID-seq7:  5’-ccagatatttcgccgcgac-3’ pID-seq8: 5’-cggggccagcaaaaaatcca-3’ pID-seq9: 5’-ccctgatttttcaccacccc-3’ pID-seq10: 5’-cttccgaaaatgcaacgcga-3’ Biobrick frt’-cm-frt’ frt’-cm-frt’-for: 5’-gaagttcctatactttttagagaataggaacttcgttgatcgggcacgtaagagg-3’ frt’-cm-frt’-rev : 5’-gaagttcctattctctaaaaagtataggaacttcttattacgccccgccctgcc-3’ In green is the frt’ sequence which is not homologous with the chloramphenicol gene. topo-frt’-cm-frt’-for: 5'-tccggcaaaaaagggcaaggtgtcaccaccctgcccttttcgccagtgtgctggatttc-3' topo-frt’-cm-frt’-rev: 5’-tgcagcggccgctactagtactctagaagcggccgcgaatgatggatatctgcagatttc-3’ In red is the iGEM restriction sites (prefix and suffix) and in green is the mutation made to suppress the EcoRI restriction site (in the Topo® XL PCR plasmid). Biobrick frt’-cm-frt’ sequencing (primer M13 as described in Topo® XL PCR Kit) FRT’-cm-FRT’-SEQ-FOR: 5’-Gaggaaacagctatga-3’ FRT’-cm-FRT’-SEQ-REV:  5’-gaccggcagcaaatg-3’