Label IT® µArray® Dual Labeling Kit
Optimized labeling kit designed for dual-channel microarray applications using Biotin and Fluorescein
- One-step Chemical Method—Easily and consistently achieve optimal 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 Biotin and Fluorescein dyes for expression profiling analysis.
Kit Components:
Each Kit contains Label IT Biotin and Fluorescein Reagents, Reconstitution Solution, 10X Labeling Buffer M, Reagent D1, Buffer N1, 0.5M EDTA, 10X STOP Reagent and 5X Fragmentation Buffer. Biotin and Fluorescein detection reagents are not included.
Data
Figure 1: The Label IT µArray Dual Kit Labels any Nucleic Acid Sample.
Figure 2: Direct mRNA Labeling for Gene Expression Analysis.
Figure 3: Sensitive mRNA Detection: < 10 Copies Per Cell.
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mRNA on Duplicate cDNA Arrays
cDNA on Duplicate oligo arrays
cRNA on Duplicate oligo arrays
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| Figure 1. The Label IT µArray Dual Kit Labels any Nucleic Acid Sample. The Label IT µArray Dual Labeling Kit was used to label mRNA directly for hybridization to duplicate double-stranded cDNA arrays (top), or cDNA and cRNA samples for hybridization to sense-strand oligo arrays (middle, bottom). For each microarray hybridization, the mRNA sample was divided, labeled independently with biotin and fluorescein, pooled, and hybridized to the indicated arrays. All microarray features should be yellow since they were derived from the same sample. Labeled samples were detected using a Streptavidin-Cy™3 and an anti-FL-Cy™5 conjugate. |
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| Figure 2. Direct mRNA Labeling for Gene Expression Analysis. To compare the effect of labeling biases on microarray data, brain and liver mRNA samples were each directly labeled for analysis in each channel and pooled appropriately for a "label-swap" experiment using spotted cDNA microarrays (top). Equivalent gene expression ratios were determined for each gene from both analyses (bottom), indicating the reproducibility of gene expression results and the absence of label incorporation bias. In the label-swap experiment, the average log2 ratio of the Cy™5/Cy™3 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. Each microarray hybridization contained six replicate features per gene and was conducted in duplicate. |
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| Figure 3. Sensitive mRNA Detection: < 10 Copies Per Cell. Sensitivity of detection with the Label IT µArray Dual Labeling Kit was determined from spiking experiments in which a specific quantity of an Arabidopsis mRNA sample was added to HeLa mRNA before labeling. An estimated mRNA copy number of ~10 copies per cell (30 pg of the spiked mRNA in 1 µg HeLa mRNA) can be detected in each channel above non-specific hybridization signal (negative controls). Fluorescent signal detected for the spiked mRNA and negative controls is corrected by subtracting the median local background value; error bars represent one standard deviation. |
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