Expression Profiling Array Services

Description
CapitalBio´s Expression Profiling Array Services guarantee the quality of your results. The exceptional quality of our services were highlighted in the recent FDA-sponsored MicroArray Quality Control (MAQC) project (Nature Biotechnology,24(9): 1140-1150, 2006). CapitalBio’s highly trained specialists manufacture, hybridize, scan, and conduct a preliminary analysis of your arrays. There is no need for you to train your lab personnel in highly specialized techniques or purchase expensive instruments to fabricate arrays in-house. Moreover, because CapitalBio´s products range from ready-to-use arrays to customizable array designs, you will be able to explore the complex bio-world and answer your research questions more quickly and cost effectively.

Key Features
CapitalBio´s pre-spotted high-density oligonucleotide microarrays consist of 70mers with sequences designed and tested to provide maximum sensitivity and specificity, while minimizing background and non-specific hybridization. Each probe is arrayed and covalently linked to the slides using optimal glass surface chemistry.
★ Highly sensitive: comparable to cDNA probes without the drawbacks of PCR amplification
★ Excellent specificity: sequence optimized to minimize cross hybridization, allowing analysis of overlapping and homologous genes
★ Normalized hybridization conditions: melting-temperature normalized oligo sets
★ Reliable and reproducible results

High Quality Array Results
Array Reproducibility


Array Sensitivity
★ Only 10-20 μg of total RNA per sample required for standard services, and requiring as little as 0.01μg of total RNA for whole genome expression profiling.
★ Absolute Sensitivity: able to detect 20pg mRNA in 10 μg total RNA.

Stringent Quality Control
Our standard services include QC checkpoints that allow our staff members to decide whether to proceed with the next step of the testing process, thereby saving expensive arrays and facilitating the interpretation of array results.
★ Visual Inspection: We validate every production lot of arrays using a visual-light scan of randomly sampled slides, allowing the identification of missed or inadequately deposited samples.
★ Batch Quality Control: Every array in a batch and every package of printed arrays are closely scrutinized before use to detect inadequate sample deposition, cross-linking, surface chemistry anomalies and oligo concentration variations outside of normal limits. This is accomplished through a variety of imaging modalities and assays using randomly selected slides from each print run. The data are compiled and included into the QC report.
★ Sample Quality Control: RNA samples are run on RNA grade agarose gels to determine if possible degradation has occurred. The concentration and OD 260/280 ratios are determined on a spectrophotometer upon reception of customer test samples. We use brand reagent kits to incorporate fluorescent dyes into cDNAs. After post-labeling purification, the labeled cDNA sample concentration, fluorescent intensity and labeling efficiency are determined, and test-reference sample normalization is conducted before applying onto arrays. These data are provided in the QC report.

Services Description
RNA Sample Preparation
Extraction and purification of RNA from tissues, cell cultures and other materials. Typically Qiagen or MACHEREY-NAGEL kits will be used for these procedures.

Initial Sample QC
RNA samples are run on RNA grade agarose gels to determine possible degradation. The concentration and OD 260/280 ratios are measured on a spectrophotometer. Customer will be notified of problematic samples. Samples unlikely to provide quality data are flagged with a recommendation that they will not be used for array hybridization. 

cDNA Synthesis (reverse transcription of RNA)
RNA samples are reverse-transcribed with submicro brand kit reagents. If sufficient sample is available, cDNA samples are tested by electrophoresis to determine the quality of synthesis, and are also checked on spectrophotometer to assure appropriate fluorescent incorporation. Corrective actions are recommended to the customer for cDNA samples that fail to meet acceptable performance criteria.

Scaled-up cDNA Syntheses
Multiple cDNA syntheses using the same RNA sample labeled with the same fluorescent dye for hybridization to additional arrays.

cDNA Testing on Experimental Arrays
cDNA samples are run on third party commercial arrays provided by the customer or CapitalBio. Arrays are hybridized according to CapitalBio´s submicro protocols. Arrays may be run with two samples simultaneously, with each sample labeled with a spectrally distinct fluorescent dye. Arrays are washed, centrifuged dry under low light conditions and then scanned.

Raw Data Generation
Image TIF files are generated. Statistical data and differential analysis files are generated using CapitalBio´s confocal scanner LuxScan 10K-A and SpotDada Pro software and transferred to Excel worksheets. All data is saved to writable CDs, which are made available to the customer for further analysis.

Ordering Information

Important Disclaimer: Microarray results will vary depending on the quality of the customer´s sample(s) and of the microarrays used for testing. During the normal course of performing array hybridizations, some experiments may fail to produce usable data, resulting in loss of test samples and customer microarrays.CapitalBio assumes no liability for the loss of customer samples or of microarrays during normal testing.CapitalBio provides no warranty as to the quality of any data generated, and shall not be held liable for any damage or cost incurred from the use of such data. In addition, the customer agrees to indemnify CapitalBio from all actions and liabilities resulted from CapitalBio´s testing of any samples or the use of any data produced by CapitalBio´s testing service.