Stratagenes Human cDNA Microarrays: Perform Gene Expression Profiling and,,,Discover New Genes ( n the Stratagene website. ...)

Stratagenes Human cDNA Microarrays: Perform Gene Expression Profiling and,,,Discover New Genes

n the Stratagene website.

Several methods are available to identify and evaluate the clones in the discovery clone collection. The collection can be searched for a specific clone by using a gene name, accession or UniGene number, nucleotide sequence, or location on a discovery-3 microarray. Additional information on the Virtual Lab website includes gel images of restriction enzyme digestions of individual clones and gel images demonstrating the length and purity of PCR products used for microarray spotting (Figure 1). Sequence information is available upon purchase of any clone.

Fig.1

Insert-Specific Priming for High-Quality Arrays

Most cDNA microarrays are created by spotting small amounts of PCR products obtained from plasmid cDNA templates onto glass microscope slides. Such PCR products are typically generated using two vector-specific primers that anneal to priming sites flanking the cDNA insert. There have been many reports that the DNA spotted onto such microarrays is often a mixture of more than one clone or an incorrect clone.

Stratagene has introduced several quality-control steps to essentially eliminate the spotting of more than one clone per spot or spotting an incorrect clone on the discovery-3 microarrays. These steps include digestion of each plasmid cDNA template with restriction enzymes to determine the DNA purity and nucleotide sequencing from the 3 end (Figure 2). The 3 sequence information is then used to design an insert-specific primer that is capable of amplifying approximately 350 bases of the 3 end of the cDNA, including the polyA tail. After PCR with the insert-specific primer, the presence of a single PCR product of the correct length is confirm
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( n the Stratagene website. ...)Stratagenes Human cDNA Microarrays: Perform Gene Expression Profiling and,,,Discover New Genes

Fig.1

Insert-Specific Priming for High-Quality Arrays