DNA Methylation Pyrosequencing Assay Is Applicable for the Assessment of Epigenetic Active Environmental or Clinical Relevant Chemicals

Table 2

Self-designed pyrosequencing assays using PSQ assay design are shown. Primer score represents the probable quality of the primer combinations (min. 0->max. 100, color score: blue > yellow > orange > red); F1: forward primer; R1: reverse primer (biotinylated); S1: sequencing primer (LINE1_CG7-14).

(a)  

Primer set 1	Score: 81
General warnings

(b)  

⇀	F1	GGTTTATTTTATTAGGGAGTGTTA
⥞	R1	AAAAAAAAACTCCCTAACC
→	S1	AGGGAGTGTTAGATAGTGG

(c)  

	PCR product	Forward PCR primer, F1	Reverse PCR primer, R1	Sequencing primer, S1
Length, bp	127	24	19	19
Position, 5′-3′		105-128	231-213	118-136
Warnings
Tm, °C		61.9	59.3	50.8
% GC	37.0	29.2	31.6	47.4
Sequence to analyze	GYGTAGGTTA TTGTGTGYGY GTATYGTGYG YGAGTYGAAG TAGGGYGAGG TATTGTTTTA TTTGGGAAG