Figure 2 (A) 異なるbatchのUniversal Human Reference RNAをCy3、Cy5でラベルし、12000スポットのヒトマイクロアレイにハイブリダイズさせた結果のスキャッタープロット。(B)同一batchのUniversal Human Reference RNAをCy3、Cy5でラベルして12000スポットのヒトマイクロアレイにハイブリダイズさせた結果の、self-to-selfスキャッタープロット。いずれのスキャッタープロットも非常によく収束しており、Universal Human Reference RNAのbatch間のばらつきがほとんど無いことが示されました。
Figure 3 Universal Human Reference RNAのAgilent 2100 バイオアナライザーでの解析結果。
Figure 4. Design of a Two-Hydribdization Experiment with Multiple Samples Using Universal Human Reference RNA
Table 2. Data Summary of the Universal Human Reference RNA on the U133A Plus 2.0 Affymetrix GeneChip
Figure 5 Total RNA isolated from 10 individual human cell lines were reverse-transcribed to cDNA, labeled with Cy5 and co-hybridized with Cy3-labeled UHRR onto 43,000-spot cDNA microarrays (Stanford University). The data was analyzed using GeneTraffic software. Approximately 6000-8000 spots out of 43,000 (14-18 %) were flagged on each microarray and excluded from further analysis. Spots with hybridization signals in Cy5 channel higher than 1000 and with Cy5/Cy3 ratio greater than 2 were collected and the number of spots with these characteristics on only one microarray was determined.
Figure 6. Self-Self Scatter Plot of the UHRR hybridized to two Affymetrix GeneChip U133A 2.0 Plus Microarray on separate occasions.
Figure 1. Cell Line Composition of the Universal Human, Mouse and Rat Reference RNAs
Table 1 Coverage of the Universal Human, Mouse and Rat Reference RNAs on Several Microarray Platforms