Description

Discover chips™ offer the possibility to study 380 genes, among the most studied of four important species: Arabidopsis, Human, Mouse, Rat.
Discover chips™ are for research use only.
These Oligonucleotide microarrays contain 380 genes selected from the most important cellular functions, allowing obtention of transcriptional and physiological informations.
The 70-mer oligos were synthesized, purified and printed in duplicate on slide (class 100 microarray cleanroom).
Four negative controls are included on the slide. Oligos controls are conceived to be unique by computing analysis via BLAST, targeting the public data bases sequences to avoid "cross-hybridization". They allow Cy3 and Cy5 signals normalization as well as normalization and control of microarrays experiments.
 

Reference	Designation
DCA	Discover™ Chip - Arabidopsis
DCH	Discover™ Chip - Human
DCM	Discover™ Chip - Mouse
DCR	Discover™ Chip - Rat

Specifications

Major functional groups spotted on the slide:

• Antigene • Apoptosis • Cell Adhesion • Cell Cycle • Transporters • Cytokine • Cytosqueleton

• Growth factors • Immunology • Kinase • Oncogene • Protease • Receptor • Transcription Factor

All oligos have a gene description, a symbol, a locus link and a GenBank code referenced in appendix. The oligo sequence is also available.

Discover Chips microarrays are produced using high quality facilities. Experiment success will depend on every protocol step and so, on target quality. You must use the protocol included in your kit for best results.

Protocol

Hybridization Protocol for DiscoverChips^TM

 1. Wash the Discover Chips to remove excess probe sequences and prepare the chip for blocking.

- 5 minutes in 2X SSC, 0.1% SDS
- 5 minutes in 2X SSC
- 5 minutes in 0.1X SSC

Transfer the microarrays to dH2O for 30 sec and then to 100% ethanol for 10 sec. Spin the microarray dry for 10 sec.
The use of High Speed microarray Centrifuge is required.

2. Blocking : microarray surface preparation to reduce background fluorescence.

3. Target prepapration and hybridization.

During this step, labeling may be performed using reverse transcriptase to generate labeled cDNA. 1 μg of labelling cDNA is suspended in 10 μl of HybIt hybridization Buffer and then hybridized at 65°c for 3 hours.
For your information:
Target purification kit allows increasing target quality by removing excess of salt, enzymes and primers.
Labelling reagents are also available. Needed volumes are indicated according to the type of target.

4. Wash the microarray to remove unbound target.

Three washes are required. Washes can be made in solutions containing PBS or SDS and a detergent decreasing gradient (e.g. Triton).
Use of ArrayIt Wash Buffer is advised. Prior to washing, remove the cover slide.
Place the slide in the first wash buffer (e.g. Wash Buffer A) for 5 minutes under agitation. Transfer the slide in the second wash buffer (e.g. Wash Buffer B) for 5 minutes.
5 minutes in the third wash solution (e.g. Wash Buffer C) should be sufficient to complete the wash process.
Wash protocol can be optimized by changing temperature and using different washes buffers.

5. Detect signals by scanning.

6. Quantify signals.

7. Perform data analysis.

ArrayIt^® Discover Chips™ microarrays are manufactured and packaged in state-of-the-art class 100 Cleanroom facilities and are stable at room temperature (25°C) for 6 months from the date of receipt. Store in the white substrate slide box and avoid exposure to elevated temperature, humidity, and dust.