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Label IT® µArray Labeling Kits, Cy™3 and/or Cy™5

Optimized labeling kit designed for dual-channel microarray applications using Cy™3 and/or Cy™5 dyes

• One-step Chemical Method—Easily and consistently achieve optimized labeling densities for microarray applications.
• Sensitive Detection—Confidently detect rare transcripts and small changes in gene expression.
• Versatile Labeling—Suitable for labeling mRNA, cDNA and cRNA templates with Cy™3 and Cy™5 dyes for expression profiling analysis.

Kit Components:

Each Kit contains Label IT Cy™3 and/or Cy™5 Reagents, Reconstitution Solution, 10X Labeling Buffer M, Reagent D1, Buffer N1, 0.5M EDTA, 10X STOP Reagent, and 5X Fragmentation Buffer.

Data

Figure 1: The Label IT® µArray Labeling Kits, Cy™3 and/or Cy™5.
Figure 2: Direct Labeling and Hybridization of mRNA Samples Using the Label IT µArray Cy™3/Cy™5 Kit.
Figure 3: Direct mRNA Labeling for Gene Expression Profile Analysis.
Figure 4: Array Based Comparative Genomic Hybridization Applications Using the Label IT µArray Cy™3/Cy™5 Kit.

Figure 1: The Label IT® µArray Labeling Kits, Cy™3 and/or Cy™5. The Label IT Cy3 and Cy5 reagents are composed of three regions: the Cy3 or Cy5 label (green), the linker (yellow) which facilitates electrostatic interactions with nucleic acids and the reactive alkylating group (blue) that covalently attaches the Label IT reagents to any reactive heteroatom within cDNA or rRNA molecules. The covalent attachment of the Label IT reagents to cDNA or cRNA does not alter their structure or downstream hybridization performance, and thus, once labeled, the Cy3 and Cy5 labeled cDNA or cRNA samples can be pooled and hybridized to gene expression microarrays. Hybridized molecules are then detected directly and analyzed to determine the gene expression profiles.

Figure 2. Direct Labeling and Hybridization of mRNA Samples Using the Label IT µArray® Labeling Kit, Cy™3/Cy™5. RNA samples were enriched for mRNA, labeled with the Label IT µArray® Labeling Kit, Cy™3/Cy™5, pooled, and hybridized to a cDNA IntelliGene™ TestARRAY v4.0 (TaKaRa Mirus Bio Inc.).

Figure 2. Direct mRNA Labeling for Gene Expression Profile Analysis. HeLa and A549 mRNA samples were labeled in a “dye-swap” experiment comparing the Label IT µArray® Labeling Kit, Cy™3/Cy™5 to enzymatic incorporation of Cy™3 and Cy™5 (SuperScript™ Direct cDNA Labeling System; Invitrogen). In this “dye-swap” experiment, the average log2(Cy™3/Cy™5) signal ratio for each gene is expected to change sign (e.g. from +1 to -1) when samples are labeled for detection in the opposite channel.

Figure 3. Array Based Comparative Genomic Hybridization Applications Using the Label IT µArray® Labeling Kit, Cy™3/Cy™5. Mse I/Alu I digested human male and female DNA samples were labeled with Cy™5 and Cy™3, respectively, using the kit. 2.5 µg of each sample was hybridized to a BAC array (Spectral Genomics), according to the recommended procedure for CGH analysis. The red features indicate signal from the male sample; green features indicate signal from the female sample.