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. 2014 May 27;133(8):1059–1073. doi: 10.1007/s00439-014-1447-z

Table 1.

List of primers and probes used in this study

Method Primer sequence
Pyrosequencing
 CpG island FP: 5ʹ-TGTGTTAAGTTGTTTAGGGGTTTTA-3ʹ
RP: 5ʹ-ACTACAATCTCTACCTCCCAAATTC-biotin-3ʹ
SP: 5ʹ-AATATTATGTGGGAAATTGT-3ʹ
 5ʹUTR FP: 5ʹ-TTGGAGTATATAGGTAGTATTA-3ʹ
RP: 5ʹ-TCCCCTAAAATCTCTAAC-biotin-3ʹ
SP: 5ʹ-TTAGTTGTGTAGGAGAAAGG-3ʹ
GDF5 expression FP: 5ʹ-CTGTGATTCCAGGAGTGCAG-3ʹ
RP: 5ʹ-ATCCTCTTCATTGACTCTGCC-3ʹ
Pr: 5ʹ-CCACGACCATGTCCTC-3ʹ
rs143383 genotyping FP: 5ʹ-AGTCAGTTGTGCAGGAGAAAGG-3ʹ
RP: 5ʹ-GCAGCTGAAAATAACTCGTTCTTGAA-3ʹ
Pr: 5ʹ-AGAAAGCCACCGCC-3ʹ
DY-682 EMSA probes
 rs143383 C allele unmethylated FP: 5ʹ-GAGAAAGGGGGCGGTCGGCTTTCTCC-3ʹ
RP: 5ʹ-GGAGAAAGCCGACCGCCCCCTTTCTC-3ʹ
 rs143383 C allele methylated FP: 5ʹ-GAGAAAGGGGGCmeGGTCGGCTTTCTCC-3ʹ
RP: 5ʹ-GGAGAAAGCCGACCmeGCCCCCTTTCTC-3ʹ
 rs143383 T allele unmethylated FP: 5ʹ-GAGAAAGGGGGCGGTTGGCTTTCTCC-3ʹ
RP: 5ʹ-GGAGAAAGCCAACCGCCCCCTTTCTC-3ʹ
 rs143383 T allele methylated FP: 5ʹ-GAGAAAGGGGGCmeGGTTGGCTTTCTCC-3ʹ
RP: 5ʹ-GGAGAAAGCCAACCmeGCCCCCTTTCTC-3ʹ
SP1/SP3 consensus competitor FP: 5ʹ-AATTGGGGGGGCGGGGGTACGTAGCA-3ʹ
RP: 5ʹ-TGCTACGTACCCCCGCCCCCCCAATT-3ʹ
Cloning into pGL3-enhancer FP: 5ʹ-GGGGACTAGTGGATTCAAAACTAGGGGG-3ʹ
RP: 5ʹ-GGGGAAGCTTCCGCTGAATGACACCAAAG-3ʹ
Cloning into EGFP-N1
 SP1 FP: 5ʹ-GGGGGAATTCATGGATGAAATGACAGCTGTG-3ʹ
RP: 5ʹ-GGGGCCGCGGGAAGCCATTGCCACTGATATT-3ʹ
 SP3 FP: 5ʹ-GGGGGAATTCATGACCGCTCCCGAAAAGCCC-3ʹ
RP: 5ʹ-GGGGCCGCGGCTCCATTGTCTCATTTCCAGA-3ʹ

The bold and underlined bases in the EMSA probes indicate the position of rs143383. The series of underlined bases in the SP1/SP3 competitors indicate the site of the consensus sequence for SP1/SP3

FP forward primer, RP reverse primer, SP sequencing primer, Pr probe, me methylated