IGEM:MIT/2005/Primers
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The melting temperatures were calculated mostly by the following site: Oligonucleotide Primer Check
- scFv (pCT-4-4-20, pCT302 S101A, pCT-4M5.3 anti-fluorescein plasmids)
FWD: 5' CATCAT GAATTC GCGGCCGC T TCTAGA GACGTCGTTATGACTCAAACACC 3' T_m,i=71.8 T_m,f=88.5 rand EcoRI NotI XbaI Primer
REV: 5' GCTCAT ACTAGT TTATTA CTTATCATCATCATCCTTATAATC GGAGACGGTGACTGAGGTTCC 3' T_m,i=74.7 T_m,f=87.1 rand SpeI Stop FLAG Tag Primer
- FecA
fecA-Fwd 5’CATCAT GAATTC GCGGCCGC TCTAGA TGACGCCGTTACGCGTTTTTCG 3’ T_m,i=73.2 T_m,f=89.6 rand EcoRI NotI XbaI primer fecA-Rev 5’GCCATA ACTAGT TTATTA TCAGAACTTCAACGACCCCT 3’ T_m,i=68.2 T_m,f=79.9 rand SpeI stop primer
Middle Fwd- 5'GGACGGCAGCGGACTGCAAGTAAAACCGCTGGGAAATAAC 3' T_m=84.7 Middle Rev- 5’GTTATTTCCCAGCGGTTTTACTTGCAGTCCGCTGCCGTCC 3' T_m=84.7
fecA Promoter Fwd- 5’CATCAT GAATTC GCGGCCGC T TCTAGA TGGATAAACATTTCACCACTG 3' T_m,i=65.0 T_m,f=86.2 rand EcoRI NotI XbaI primer fecA Promoter Rev- 5'GCCATA CTGCAG CGGCCGC T ACTAGT CACAAGTCGATACTCAGCTTG 3' T_m,i=68.9 T_m,f=89.8 rand PstI NotI SpeI primer
- FecR
fecR-Fwd 5' CATCAT GAATTC GCGGCCGC TCTAGA tgaatcctttgttaaccgattccc 3'T_m 66 rand EcoRI NotI XbaI primer fecR-Rev 5' GCCATA ACTAGT TTATTA ttacagtggtgaaatgtttatccag 3' T_m 68 rand SpeI stop primer Middle Fwd 5'gtgaaagcctacagttcagcgcctcag3' T_m 84 Middle Rev 5'ctgaggcgctgaactgtaggctttcac3' T_m 84
- FecI
fecI-Fwd 5'CATCAT GAATTC GCGGCCGC T TCTAGA tgtctgaccgcgccactacc3' T_m 68 rand EcoRI NotI XbaI primer fecI-Rev 5'GCCATA CTGCAG CGGCCGC T ACTAGT A TTATTA tcataacccatactccagacggaa3' T_m 70 rand PstI NotI SpeI stop primer
- MalE (updated/fixed)
fwd: 5' CCGTTA GAATTC GCGGCCGC T TCTAGA atgaaaataaaaacaggtgcacgc3' T_m,i=68.8 T_m,f=85.5 junk EcoRI NotI XbaI Primer * rev: 5' ACTGAG CTGCAG GCGGCCGC T ACTAGT TTATTA TTACTTGGTGATACGAGTCTGC 3' T_m,i=69.5 T_m,f=89.4 junk PstI NotI SpeI Stop Primer
- PhoA
fwd: 5' CATGAG GAATTC GCGGCCGC T TCTAGA ATGAAACAGAGCACCATTGCG 3' T_m,i=68.9 T_m,f=88.8 junk EcoRI NotI XbaI Primer rev: 5' AAGCTA CTGCAG GCGGCCGC T ACTAGT TTATTA TTATTTCAGGCCCAGCGCC 3' T_m,i=69.7 T_m,f=89.8 junk PstI NotI SpeI Stop Primer
Middle(1) fwd: 5' caccgcggaactgcaagatgcgaccccggcg 3' T_m=88.8 * rev: 5' CGCCGGGGTCGCATCTTGCAGTTCCGCGGTG 3' T_m=88.8 *
Middle(2) fwd: 5' aggcttttttctgcaagtggaaggcgcgagc 3' T_m=82.2 * rev: 5' GCTCGCGCCTTCCACTTGCAGAAAAAAGCCT 3' T_m=82.2 *