Identification of methylation profile of HOX genes in extrahepatic cholangiocarcinoma

Table 1 Primer sequences, fragments and annealing temperature used for bisulfite-polymerase chain reaction and reverse transcription-polymerase chain reaction

Genes	Primer 5'→3'	Annealing temperature (°C)	Size (bp)
BSP
HOXA2	L: TGTTTTAATAGAATTTATGTGGTTGG	56	190
	R: ATAACTACCCTCTACCTCCCCC
HOXA5	L: GTTTTGGAGAAATATTATATAAAAGTTATT	50	145
	R: CAATTAAAAAAATAAATCCTACCC
HOXA11	L: TGAGTATAAGTATGTTGTATGGGGG	60	148
	R: TTATAACCACCTCAAAAAAAACAAC
HOXB4	L: TAGAGGTGAGGTAGAATAGGAGGGT	60	204
	R: ACCCAACACCAAAATTTACATAAAA
HOXD13	L: GTGGGTTTAGTTAGGTTTGGGT	60	239
	R: TCTAACCCTCTCTCCCTCTATAAAC
RT-PCR
HOXA2	F: GTCACTCTTTGAGCAAGCCC	59	345
	R: TAGGCCAGCTCCACAGTTCT
HOXA5	F: ACTCCGGCAGGTACGGCTACG	62	259
	R: -CCGCTGGAGTTGCTTAGGGAG
HOXA11	F: TGCCAAGTTGTACTTACTACGTC	61	181
	R: GTTGGAGGAGTAGGAGTATGTA
HOXB4	F: GTGCAAAGAGCCCGTCGTCTA CC	65	161
	R: CGTGTCAGGTAGCGGTTGTAGTG
HOXD13	F: TGCTCCTCTTCTGCCGTTGT	55	467
	R: CCTGTGGCTGGTCCTTGGT

BSP: Bisulfite-polymerase chain reaction; RT-PCR: Reverse transcription-polymerase chain reaction.