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Table 1 Primers and amplification conditions for PCR, pyrosequencing, and RT-qPCR experiments

From: DNA methylation of the allergy regulatory gene interferon gamma varies by age, sex, and tissue type in asthmatics

Gene/region Assay Primers PCR Conditions
IFNγ promoter CpG -186 PCR and Pyrosequencing F: 5’-biotin-AGATGGTGATAGATAGGTAGGGATGATA-3’ 95°C, 15 min; 45 cycles of 95°C, 30 sec; 55°C, 30 sec; 72°C, 30 sec; 72°C, 10 min; 4°C hold
R: 5’-TCCCACCAAAATAACACAAATAAACAT -3’
S: 5’-AAATAAACATAATAAATCTATCTCA-3’
IFNγ promoter CpG -54 F: 5’ATGTGTTGTATTTTTTTTGGTTGTTGGTAT-3’
R: 5’-biotin-TATCATCCCTACCTATCTATCACCATCTC-3’
S: 5’-ATTGAAGTTTTTTGAGGATT-3’
IFNγ promoter (target) RT-qPCR F: 5’-TCGGTAACTGACTTGAATGTCCA-3’ 95°C, 3 min; 40 cycles of 90°C, 10 sec; 55°C, 30 sec; 4°C hold
R: 5’-TCGCTTCCCTGTTTTAGCTGC-3’
GAPDH (internal control) F: 5’-ACAACTTTGGTATCGTGGAAGG-3’
R: 5’-GCCATCACGCCACAGTTTC-3’
  1. F (forward primer), R (reverse primer), S (sequencing primer), IFNγ (interferon gamma), GAPDH (glyceraldehyde 3-phosphate dehydrogenase), min (minutes), sec (seconds).