Table 1.

PCR primers and conditions used to determine the methylation status of promoters of the PTEN gene and PTENP1 pseudogene

Primer	Nucleotide sequence, 5’->3’	Ta., °C	Number of PCR cycles	PCR product size, b.p.
PNP1-U-F	TTGTAGTTGTGATGGAAGTTTGAAT	64	33	156
PNP1-U-R	CCACCCCCACAAATACTCACA
PNP1-M-F	TGTAGTCGTGATGGAAGTTTGAAT	63	33	152
PNP1-M-R	CCCCCGCGAATACTCACG
PN2-U-F	TTGTAGTTATGATGGAAGTTTGAG	61	33	165
PN2-U-R	CCACCACCACAAACCAACCA
PN2-M-F	TTGTAGTTATGATGGAAGTTTGAG	61	33	162
PN2-M-R	CGCCGCAAACCGACCGA
PN4-U-F	GTTGGGGTGTGTGGAGTTTGGTT	61	36	135
PN4-U-R	CCCTCAAACTCCAAATCAATTCACAA
PN4-M-F	CGCGCGGAGTTTGGTTTCG	62	32	117
PN4-M-R	CAAATCGATTCGCGACGTCG
PN5-U-F	TATTAGTTTGGGGATTTTTTTTTTGT	60	36	186
PN5-U-R	CCCAACCCTTCCTACACCACA
PN5-M-F	GTTTGGGGATTTTTTTTTCGC	60	36	178
PN5-M-R	AACCCTTCCTACGCCGCG

Note. M – methylated; U – unmethylated; F – a forward primer; R – a reverse primer; PNP – primers for amplification of pseudogene regions; PN – primers for amplification of PTEN gene regions.