WHAT DO YOU HAVE TO DO?
Prepare your total RNA samples from tissues of interest and send it on dry ice to us. Our Hybridization service starts with as little as 100 ng of total RNA.

Depending on the type of experiment, we label each cDNA with the same dye (Cy3) and hybridize them on individual arrays (single channel experiments). Alternatively, we do co-hybridizations: We label RNA from control tissue and from the test tissue with Cy3 and Cy5, repectively and hybridize them both on one slide.

In order to have all information necessary, please send us the Hybridization Service Order form, together with your samples.

WHAT CAN WE DO FOR YOU?

INITIAL CONTROL

Upon arrival, we verify the content of your package carefully. All samples are transferred immediately to –80°C. An aliquot is tested with Agilent 2100 bioanalyzer system that requires only minimum amounts of your precious RNA samples. We will send you a feedback report of these initial controls immediately.

LABELING

Labeling is performed incorporating Cy3 or Cy3 and Cy5. Depending on the available amounts of total RNA, you can choose from different labeling protocols.

For more information click here.

After labeling, we monitor quality, yield, and incorporation rates for every sample.

HYBRIDIZATION

Hybridizations are performed in a state-of-the-art hybridization station. The stringency of  hybridization condition is adjusted carefully to ensure highest signal intensity at the lowest background contamination. An excellent signal-to-noise ratio is also supported by our proprietary algorithms for designing the oligos used on our catalog and custom arrays.

For more information click here.

Additionally, our workflow is highly automated in order to get the best reproducibility during this crucial step.

SCANNING

Every channel (Cy3, Cy5) is scanned three times with increasing photomultiplier gain settings ensuring coverage of the full dynamic range. We are using a state-of-the-art scanner at 10 µm resolution.

DATA ANALYSIS

Unique pixel selection algorithms are used to determine signal intensities for every single spot. Intensity values are further processed using our proprietary software MAVI, which solves saturation problems and thus, enlarges the dynamic range. This is achieved by linear regression analysis of the intensity data gained from images of the three scans. The MAVI result files are exported into individual Excel sheets for each slide. These Excel sheets combine array with gene description data and contain all raw data, normalized and background corrected intensities as well as ratio values, calculated according to your requirements.

Our proprietary software Genowiz™ is used to visualize this data, e.g., creating histograms and scatter plots. Additionally, if dye swap experiments are performed, average of both slides is calculated. Further analyses such as clustering or statistics are available on request.

WHAT DO YOU RECEIVE?

Compact Disk with all results and the complete analysis which includes:

1. Control sheet protocolling every step, according to ISO 9001
2. Results of evaluation of the total RNA, evaluation of the labeled samples (dye incorporation efficiencies)
3. TIFF images from scanning, overlay image, MAVI data
4. Excel sheets combining array with gene description data, histogram and scatter plots
• Hybridized arrays in light protected packaging
• QC sheet protocolling initial control