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Table 2 Characterization of MSP primers (M – methylated; U – unmethylated) used in the study. The underlined nucleotides in forward and reverse primers indicate the presence of methylated cytosines in DNA sequence

From: Quantitative analysis of mRNA expression levels and DNA methylation profiles of three neighboring genes: FUS1, NPRL2/G21 and RASSF1A in non-small cell lung cancer patients

Gene

Forward primer

Primer position

Reverse primer

Primer position

Product length [bp]

Annealing temp. [°C]

FUS1

M: TGTTATCGTGGATTAGATATTGTTC

−559 to −482

M: ACTATATTTTTACGATTACCACGCT

−375 to −349

207

56.5

U: TTATTGTGGATTAGATATTGTTTGT

−557 to −480

U: ACTATATTTTTACAATTACCACACT

−375 to −349

205

56.5

NPRL2/G21

M: GTTCGGTTATTGTTATGGGTAGC

+108 to +131

M: AACCAATTAAACTCTCGAAAACGT

+238 to +261

153

52.5

U: GGTTTGGTTATTGTTATGGGTAGTG

+107 to +132

U: ACCAATTAAACTCTCAAAAACATC

+238 to +261

153

56.5

RASSF1A

M: ATATTTTTTCGATTTGGAGTTTTTTC

−599 to −573

M: CTACACTATAACCTACCCATCCTCG

−409 to – 384

215

56.5

U: TTTTTTTGATTTGGAGTTTTTTTGT

−599 to −573

U: TACACTATAACCTACCCATCCTCAC

−410 to −385

211

57.5