In vivo footprinting |
|
Linker adapter (equimolar amounts) |
GCGGTGACCCGGGAGATCTGAATTC (also used in LMPCR) and GAATTCAGATC |
For the non-transcribed strand |
|
Extension oligo |
CAAGCAGTCAGCAGAGGATT |
PCR and hybridization oligo |
TGGCCAGACTTTGTTTCCCAAGGGTCA |
For the transcribed strand |
|
Extension oligo |
GTAAATTACCGTTCCTCCCT |
PCR and hybridization oligo |
TCGGGTTGCGGACGCTCCA |
EMSAs |
|
Forward |
CTGTTCCGCCTGGCCCACGTGACCCGCCGAGCATAA |
Reverse |
TTATGCTCGGCGGGTCACGTGGGCCAGGCGGAACAG |
Site-directed mutagenesis |
|
pHOGH/4.5 E-box M1 |
|
M1 forward |
CTGTTCCGCCTGGCCCACGTGAtCCGCCGAGCATAA |
M1 reverse |
TTATGCTCGGCGGaTCACGTGGGCCAGGCGGAACAG |
pHOGH/4.5 E-box M2 |
|
M2 forward |
CTGTTCCGCCTGGCCCAtGTGACCCGCCGAGCATAA |
M2 reverse |
TTATGCTCGGCGGGTCACaTGGGCCAGGCGGAACAG |
pHOGH/4.5 E-box M3 |
|
M3 forward |
CTGTTCCGCCTGGCCtACGTGACCCGCCGAGCATAA |
M3 reverse |
TTATGCTCGGCGGGTCACGTaGGCCAGGCGGAACAG |
pHOGH/4.5 Δ E-box |
|
Del E-box F |
CATCAGCTGTTCCGCCTGGCCGCCGAGCATAAATGTG |
Del E-box R |
CACATTTATGCTCGGCGGCCAGGCGGAACAGCTGATG |
ChIP |
|
For the proximal E-box |
|
Forward |
TTGCAACGCCCGGCCAGAAA |
Reverse |
TCGGGTTGCGGACGCTCCA |
Nested hybridization oligo |
ATAAATGTGACCGGCCGCGGCTC |
For the internal E-box |
|
Forward |
GATGACACTGAGGCTCAGA |
Reverse |
GCTGAAATCGGTCTTGGTTGATTTCAGCC |
Nested hybridization oligo |
GGTCACACAGTAAGTTCAGCCTGCTCT |