Affymetrix GeneChip Human Genome HG-U133A [HG-U133A]

New insights on human T cell development by quantitative T cell receptor gene rearrangement studies and gene expression profiling.

rawData

To gain more insight into initiation and regulation of T cell receptor (TCR) gene rearrangement during human T cell development, we analyzed TCR gene rearrangements by quantitative PCR analysis in nine consecutive T-cell developmental stages, including CD34+ lin- cord blood cells as a reference. The same stages were used for gene expression profiling using DNA microarrays.

cell_type

Transcription profiling by array of human T-cell differentiation

"To gain more insight into initiation and regulation of T cell receptor (TCR) gene rearrangement during human T cell development, we analyzed TCR gene rearrangements by quantitative PCR analysis in nine consecutive T-cell developmental stages, including CD34+ lin- cord blood cells as a reference. The same stages were used for gene expression profiling using DNA microarrays."

To gain more insight into initiation and regulation of T cell receptor (TCR) gene rearrangement during human T cell development, we analyzed TCR gene rearrangements by quantitative PCR analysis in nine consecutive T-cell developmental stages, including CD34+ lin- cord blood cells as a reference. The same stages were used for gene expression profiling using DNA microarrays.


Affymetrix:Protocol:Hybridization-EukGE-WS2v3

Title: Affymetrix EukGE-WS2v3 Hybridization. Description:

Affymetrix:Protocol:Hybridization-FlexGE-WS2v3

Affymetrix:Protocol:Percentile

Title: Affymetrix CEL Analysis (Percentile). Description:

P-MEXP-6370

RNA was isolated using RNeasy columns (Qiagen, Hilden, Germany) according to the manufacturers protocol. RNA quality was assessed using the Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA) After this amplification was performed as described in LABELPTCL6371

P-MEXP-6371

An RNA Transcript Labeling Kit (Enzo Diagnostics, Farmingdale, NY) was used for the production of biotin-labeled cRNA targets by in vitro transcription from T7 RNA polymerase promoters, purified with Qiagen RNeasy Mini Kit (Qiagen, Chatsworth, California) and fragmented by incubation at 94 C for 35 min. 

P-MEXP-6602

For each biological repeat material obtained from 5 thymi was pooled. Median age of 6 months, range; 7 weeks to 3 years.

P-MEXP-6603

For analysis of umbilical cord blood CD34+ mononuclear cell material from 5 different donors was pooled for each biological repeat.

P-MEXP-6627

RNA was isolated using RNeasy columns (Qiagen, Hilden, Germany) according to the manufacturers protocol. RNA quality was assessed using the Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA). A first cycle of amplification was performed according to the GeneChip Eukaryotic small sample target labeling assay version II (affymetrix). After this a second round of amplification was performed as described in LABELPTCL6371

Transcription profiling by array of human T-cell differentiation

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