WT-Ovation™ One-Direct Amplification System
The WT-Ovation™ One-Direct RNA Amplification System enables whole-transcriptome gene expression analysis from a single cell. This single-day method prepares targets for microarray or qPCR analysis from lysate of one or more cells, or as little as 10 pg purified total RNA. WT-Ovation™ One-Direct is specifically designed to enable the direct interrogation of cell lysate, eliminating the necessity for inefficient and time-consuming RNA purification. This direct-from-cell-lysate capability along with NuGEN’s proven Ribo-SPIA® amplification technology enables researchers to explore true, single-cell biology for the first time.
The amplified cDNA can be used directly in qPCR analysis. Alternatively, it can be labeled using the Encore™ Biotin Module for analysis on Affymetrix GeneChip® 3', Exon 1.0 ST, and Gene 1.0 ST expression arrays. Using NuGEN's Agilent Solution or Illumina Protocol, targets from One-Direct system may be interrogated on Agilent Dual-Mode Gene Expression Arrays, or Illumina BeadArray Expression Arrays.
The WT-Ovation One-Direct RNA Amplification System provides optimized reagent mixes including NuGEN's Direct Lysis Buffer, and a protocol to process 12 samples (cat.# 3500-12).
Microarray Analysis of 10 Cell Lysate Samples Amplified With the WT-Ovation™ One-Direct System
In order to demonstrate array performance with cell lysates, we performed triplicate amplifications representing 10-cell samples of untreated THP-1 cells (red), PMA treated/serum starved cells (green), and PMA treated/serum starved cells after exposure to LPS (purple) using the WT-Ovation One-Direct System. The amplified cDNA was labeled and hybridized to Affymetrix GeneChip® Human Gene 1.0 ST arrays. Principal Components Analysis (PCA) was performed using Partek Genomics Suite software. An analysis of the array data reveals differential expression of genes consistent with published data on gene expression profiles in LPS stimulation models (not shown).
