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

Table 1

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_CG1-6).

(a)  

Primer set 1	Score: 76
General warnings

(b)   

⇀	F1	GGGAGGAGTTAAGATGGT
⥞	R1	ATAAACCCCATACCTCAAA
→	S1	GGGAGGAGTTAAGATGGT

(c)  

	PCR product	Forward PCR primer, F1	Reverse PCR primer, R1	Sequencing primer, S1
Length, bp	109	18	19	18
Position, 5′-3′		3-20	111-93	3-20
Warnings
Tm, °C		60.2	60.9	53.1
% GC	33.0	50.0	36.8	50.0
Sequence to analyze	YGAATAGGAA TAGTTTYGGT TTATAGTTTT TAGYGTGAGY GAYGTAGAAG AYGGGTGATT TTTGTATTTT TATTTGA