Human cDNA matched pair products include: Primary Pair (PP), or Primary and Metastatic Pair (PM). PP consists of cDNAs isolated from primary tumor and its adjacent normal tissue; PM consists of cDNAs from primary tumor and corresponding metastatic tumor. cDNAs in each pair are prepared from the same donor. This product line is designed for identifying tumor-specific genes and tumor.

The cDNA panel is comprised of 2 vials each containing 10ul of PCR Ready First Strand cDNA. All cDNAs in a panel are made under optimal and consistent conditions, which ensure that representation is maintained from total RNA to cDNA. The First-Strand cDNA is synthesized from RNA isolated from a wide variety of documented normal and tumor tissues. Total RNA used for cDNA synthesis is isolated by modified guanidine thiocyanate techniques.

Primary Tumor 1x10rxn Metastatic Tumor 1x10rxn

~11ug total RNA was primed by an oligo dT primer and reverse transcribed by MMLV reverse transcriptase in 40ul final volume. RT reaction was stopped by heating at 65°C for 10 minutes. 1ul cDNA is sufficient for one PCR reaction. The 5' end of human clathrin cDNA (a 6kb gene) has been amplified by PCR from all of the cDNAs.

cDNA in 1X RT buffer (50 mM Tris Cl, pH 8.3, 75mM KCl, 3mM MgCl2, 10mM DTT).

Ready to use for PCR quantification of gene expression Oligo dT primer used to ensure the entire 3' end of cDNA is present With some cDNA used as templates, 12kb PCR amplicon was obtained to ensure intact cDNAs The largest selection of cDNAs from different tissues on the market Placenta cDNA is included in every panel as an interpanel control Documentation of tissues' clinical histories available (additional cost) Economical way to get cDNA from multiple tissues

Immediate PCR amplification of known genes for their expression Quantification of low copy gene expression in multiple tissues Identification of tissue-specific expression of target genes Analysis of gene expression in rare tissues Verification of genetic mutation Comparison of a specific gene between different tissues Analysis of mRNA alternative splicing

1. The integrity of the RNA used for cDNA synthesis is examined by visual inspection for the presence of intact bands of 18s and 28s ribosomal RNA when electrophoresed on a denaturing agarose gel. The quality and purity of total RNA were tested by spectrophotometer. A260/280 is between 1.8 and 2.0 (detetcted in 10mM Tris-Cl, pH 7.5). The ratio of 28S/18S is > 1.

2. The RNA used for cDNA synthesis is treated by DNase I and is tested as DNA free RNA by PCR.

3. The synthesized cDNA was 5” selected to ensure its full length. The cDNA was used as a template for PCR amplification of B-actin gene and an 838 basepair B-actin band was visualized on 2% agarose gel. B-actin control primer is included. It is enough for 10 PCR reactions.

Store cDNA panel at -20ºC Stable for 12 months.