Reference Gene Selection

Reference Gene Selection implements Vandesompele's method to identify and validate stable housekeeping (internal control) genes and determine robust normalization factors for RT-PCR gene-expression analysis.

Key Features:

• Systematic Identification: Systematically evaluates housekeeping genes across different abundance levels and functional classes to assess candidate reference genes.
• Stability Assessment: Identifies the most stably expressed genes within specific tissue sets to minimize variability in gene-expression data.
• Normalization Strategy: Determines the minimum number of genes required to compute a reliable normalization factor using the geometric mean of multiple selected housekeeping genes.
• Validation with Microarray Data: Validates the geometric mean normalization factor and reference-gene selection by analysis of publicly available microarray data.

Scientific Applications:

• High-throughput RT-PCR expression profiling: Supports accurate quantification in high-throughput RT-PCR expression studies.
• Detection of small expression differences: Facilitates detection and analysis of subtle changes in gene expression.

Methodology:

Evaluates ten housekeeping genes across various human tissues and validates the geometric mean of multiple selected housekeeping genes as a normalization factor using analysis of publicly available microarray data.