Agilent Microarray SAMPLE SUBMISSION

Overview

Agilent microarray hybridization is carried out in Agilent Microarray Hybridization Chambers with labeled cRNA and scanned using Agilent Microarray Scanner (part number G2565AA). Microarray data is extracted with Agilent Feature Extraction Software.

Services

The user is responsible for target preparation. We recommend use of Agilent protocol for target preparation. Please refer to the protocol of one-color microarray-based gene expression analysis (P/N G4140-90040) or two-color microarray-based gene expression analysis (P/N G4140-90050) for detail information.

Our core facility will provide all materials and reagents needed for the hybridization, washing, and microarray scanning. The user purchases the array chips from Agilent directly as our core does not provide arrays for individual purchase.

 Quality Control

The high quality of RNA sample is essential to ensure the quality of microarry results. The user can measure cRNA using NanoDrop ND-1000 UV-UIS Spectrophotometer, which is located in the Research Building, E401. Acceptable A260/A280 ratios are in the range 1.8-2.2.

Acceptable cRNA yield and specific activity values are as follows:

1.      For one-color microarray-based gene expression analysis cRNA yield > 1.65 ug cRNA specific activity > 9.0 pmol Cy3 per ug cRNA

2.      For two-color microarray-based gene expression analysis cRNA yield >825 ng cRNA specific activity > 8.0 pmol Cy3 or Cy5 per ug cRNA

Sample Submission

1.      For samples prepared from one-color microarray-based gene expression analysis protocol Amount required: 2.0 ug of cyanine 3-labeled cRNA

2.      For samples prepared from two-color microarray-based gene expression analysis protocol Amount required: 1.0 ug of cyanine 3-labeled cRNA and 1.0 ug of cyanine 5-labeled cRNA