Hybridization of a DNA-Chip with 18S-rDNA PCR-products from the algae Prymnesium patelliferum

1. Positive control blank EUK 1209 blank CHLO 01 CHLO 01 CHLO 02 CHLO 02 PRYM 01 PRYM 01 blank Positive control PRYM 02 PRYM 02 PRYM 03 PRYM 03 DINO B DINO B DINO E-12 DINO E-12 BOLI 01 BOLI 01 BOLI 02 BOLI 02 Positive control blank HETERO 01 HETERO 01 PELA 01 PELA 01 EUK 1209 EUK 1209 CHLO 01 CHLO 01 CHLO 02 CHLO 02 Positive control blank Positive control blank PRYM 01 PRYM 01 PRYM 02 PRYM 02 PRYM 03 PRYM 03 Positive control blank Prymnesium patelliferum (Prymnesiophyceae) Label: Cy5 Alexandrium tamarense (Dinophyceae) Label: Cy3- DINOE-12 Hybridization of a DNA-Chip with 18S-rDNA PCR-products from the algae Prymnesium patelliferum and Alexandrium tamarense

2. Target DNA hybridized to homologous DNA on the slide Target-DNA labeled with a fluorophore Spots of DNA linked to the surface e.g. Oligonucleotides, PCR-Products, cDNA Glass-slide with a modified surface Schematic drawing of a typical Chip-Experiment

3. Target-DNA labeled with a fluorophore DINO E-12 Target DNA hybridized to homologous DNA on the slide Prymnesium patelliferum (Prymnesiophyceae) Label: Cy5 Spots of DNA linked to the surface e.g. Oligonucleotides, PCR-Products, cDNA DINO E-12 Glass-slide with a modified surface Alexandrium tamarense (Dinophyceae) Label: Cy3 On a DNA-Chip the Oligonucleotid DINO E-12 hybridizes to a Dinophyceaebut not to a Prymnesiophyceae

4. What you see on the slide • I hope the schematic drawing is self-explaining.... • We hybridized one slide with PCR-Products from 18S rDNA of Prymnesium patelliferum and Alexandrium tamarense • The PCR-Product from Prymnesium patelliferum was labeled with Cy5 • The PCR-Product from Alexandrium tamarense was labeled with Cy3 • Both PCR-Products have been hybridized simultaniously on one slide • The white spots are the positive controle for the hybridization