Promoter Methylation of RASSF1A Associates to Adult Secondary Glioblastomas and Pediatric Glioblastomas

Table 1

Primers and conditions for PCRs.

Gene	Primer sense	Primer antisense	Size	T (°C)	Cycles
PTEN
-Methylation
-U	5′-GTGTTGGTGGAGGTAGTTGTTT-3′	5′-ACCACTTAACTCTAAACCACAACCA-3′	162 bp	62°C	38
-M	5′-TTCGTTCGTCGTCGTCGTATTT-3′	5′-GCCGCTTAACTCTAAACCGCAACCG-3′	206 bp	62°C	38
MGMT
-Methylation
-U	5′-GAGAGATTTGTGTTTTGGGTTTAGTG-3′	5′-CCTTCAACCAATACAAACCAAACAA-3′	236 bp	62°C	38
-M	5′-ATTCGCGTTTCGGGTTTAGC-3′	5′-CGACCGATACAAACCGAACG-3′	227 bp	62°C	38
-Expression	5′-GGGGAAGCTGGAGCTGTCTG-3′	5′-TCTCCGAATTTCACAACCTTCA-3′	282 bp	62°C	32
RASSF1A
-Methylation
-U	5′-GAGAGTGTGTTTAGTTTTGTTTTTG-3′	5′-CCCATACTTCACTAACTTTAAACAC-3′	183 bp	56°C	38
-M	5′-GAGAGCGCGTTTAGTTTCGTTTTC-3′	5′-ACCCGTACTTCGCTAACTTTAAACG-3′	184 bp	62°C	35
-Expression	5′-TCTGTGGCGACTTCATCTGG-3′	5′-TTGGGCAGGTAAAAGGAAGT-3′	424 bp	60°C	35
p14^ARF
-Methylation
-U	5′-TTTTTGGTGTTAAAGGGTGGTGTAGT-3′	5′-CACAAAAACCCTCACTCACAACAA-3′	132 bp	62°C	35
-M	5′-GTGTTAAAGGGCGGCGTAGC-3′	5′-AAAACCCTCACTCGCGACGA-3′	122 bp	58°C	35
-Expression	5′-CCGCCGCGAGTGAGGGTTTT-3′	5′-GCACGGGTCGGGTGAGAGTGG-3′	242 bp	65°C	32
P16^INK4A
-Methylation
-U	5′-TTATTAGAGGGTGGGGTGGATTGT-3′	5′-CAACCCCAAACCACAACCATAA-3′	150 bp	62°C	35
-M	5′-TTATTAGAGGGTGGGGCGGATCGC-3′	5′-GACCCCGAACCGCGACCGTAA-3′	151 bp	65°C	35
-Expression	5′-CGCGCGTACAGATCTCTCGAA-3′	5′-CACGGGTCGGGTGAGAGTGG-3′	161 bp	68°C	35
TFR
-Expression	5′-GTCAATGTCCCAAACGTCACCAGA-3′	5′-ATTTCGGGAATGCTGAGAAAACAGACAGA-3′	298 bp	60°C	30

U: unmethylated; M: methylated.