S7804 datasheet(13/32 Pages) ETC

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Specificity of the Assay

The specificity of the CpG WIZ™ p16 Amplification Kit is shown in Figure 2.

With a complete chemical modification reaction, U primers amplify only

unmethylated DNA (154 bp, lane 1) and M primers amplify only methylated

DNA (145 bp, lane 5). W primers amplify only DNA which is not chemically

modified, or "wild type W" (142 bp, lane 9).

Figure 2: Specificity of the CpG WIZ™ p16 Amplification Kit.

M 1 2 3 4 5 6 7 8 9 10 11 12 M

Polyacrylamide gel analysis using the primers and the control DNA samples

included in the kit is shown. (Lanes 1-3: U control DNA; lanes 4-6: M control

DNA; lanes 7-9: W control DNA; lanes 10-12: Minus DNA control. Lanes 1, 4,

7, 10: U primers; lanes 2, 5, 8, 11: M primers; lanes 3, 6, 9, 12: W primers. M

lane: 100 base-pair marker.)

Experiment Setup

CpG WIZ™ Amplification Kits (S7800, S7802, and S7804) include sufficient

reagents to analyze 25 samples with appropriate controls (105 PCR reactions).

Each experiment will include chemical modification of seven DNAs: 5

experimental DNA and 2 control DNA samples, (M and U). In the subsequent

PCR reactions, each chemically modified experimental DNA sample is

amplified with each of three oligonucleotide primer sets U, M and W. The

chemically modified genomic control DNAs, U and M, are amplified with their

corresponding primer set. Untreated W genomic control DNA is amplified with

the W primer set. Lastly, a negative PCR control (i.e. no DNA) is performed for

each set of primers.

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