( The Microar...)Microarray Target Purification Kit and,,,the Microarray Target Synthesis Kit (T7)

The Microarray Target Synthesis Kit (T7) has been developed as a robust, reliable and highly reproducible tool for the preparation of large amounts of labeled cRNA when starting with only 105 106 cells for target preparation. Major features are:

• acceptance of low amounts of cDNA (obtained from 1 g total RNA)
• short reaction time of only 2 3 hours (Figure 2)
• high yields of cRNA in the range of 100 g
• compatible to a broad spectrum of hapten or fluorophore-labeled nucleotides
• cost-effective labeling due to optimized nucleotide concentrations and reaction conditions
  
  

Targets, synthesized and labeled with the Microarray Target Synthesis Kit (T7) have been tested on various commercially available DNA arrays. We have found that it is sufficient to use only one labeled nucleotide to get reliable results compared to double-labeling procedures in the majority of experiments.

Very often sample material is available in such small quantities (starting with less than 1,000 cells) that efficient target preparation requires PCR-based amplification methods. Within the Microarray Target Synthesis Kit (PCR), we have optimized a random primer-based PCR amplification procedure which allows the efficient and reproducible amplification of mRNA out of 50 ng total RNA.

The resulting PCR product can then be labeled by different methods, e.g., with the Microarray RNA Target Synthesis Kit (T7) to obtain single stranded cRNA.




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