100 ng of total RNA from PBMC from 4 controls and 5 PE at week 22–24 was subjected to GeneChip HT One-Cycle cDNA Synthesis Kit and GeneChip HT IVT Labeling Kit, following the manufacturer’s protocol for whole genome gene expression analysis (Affymetrix, Santa Clara, CA). Labeled and fragmented single stranded cDNAs were hybridized to the GeneChip Human Gene 1.0 ST Arrays (28,869 transcripts) (Affymetrix). The arrays were washed and stained using FS-450 fluidics station (Affymetrix). Signal intensities were detected by Hewlett Packard Gene Array Scanner 3000 7G (Hewlett Packard, Palo Alto, CA). The scanned images were processed using Affymetrix GeneChip Command Console (AGCC). The CEL files were imported into Partek Genomics Suite software (Partek, Inc. MO). Robust microarray analysis (RMA) was applied for normalization. Differentially expressed genes between groups were identified using one-way ANOVA analysis. Cluster analysis were generated in Partek Genomics Suite. Further bioinformatics analysis was conducted on the significant genes to identify functional significance by means of Ingenuity Pathways Analysis (Ingenuity Systems, Redwood City, CA).
Genechip ht ivt labeling kit
Manufactured by Thermo Fisher Scientific
The GeneChip HT IVT Labeling Kit is a laboratory equipment product designed for the preparation of labeled cRNA samples for gene expression analysis using microarray technology. The kit provides reagents and protocols for in vitro transcription and labeling of cRNA samples from total RNA or purified mRNA.
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Lab products found in correlation
Ingenuity pathways analysis, by Qiagen (1 mentions)
Genechip command console, by Thermo Fisher Scientific (1 mentions)
Genechip human gene 1.0 st array, by Thermo Fisher Scientific (1 mentions)
Genearray scanner 3000 7g, by Hewlett-Packard (1 mentions)
Fs 450 fluidics station, by Thermo Fisher Scientific (1 mentions)
Rna labchip kit, by Agilent Technologies (1 mentions)
Genechip 3 ivt express kit, by Thermo Fisher Scientific (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Agilent rna 6000 nano kit, by Agilent Technologies (1 mentions)
2 protocols using genechip ht ivt labeling kit
1
Microarray Analysis of PBMC in Preeclampsia
2
Transcriptome Analysis of MDCK Cells
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Total RNA was isolated from the tenth generation of MDCK cells with CPV-LZ1 infection using TRIzol reagent (Life Technologies) following the manufacture’s instruction. The uninfected F10 MDCK cells act as mock control. The RNA quality was checked using a NanoVue UV spectrophotometer (Thermo Scientific, Rockford, IL, USA). Qualified total RNA was further purified by RNA LabChip kit (Agilent Technologies, Santa Clara, CA, USA).
Samples RNA was amplified using a GeneChip 3‘IVT Express Kit (Affymetrix, Cleveland, OH, USA) to generate first strand cDNA and second strand cDNA, respectively. Then complementary RNA (cRNA) was produced and labeled using GeneChip HT IVT Labeling Kit (Affymetrix). The labeled cRNA was purified and quality test by an Agilent RNA 6000 Nano Kit (Agilent Technologies), followed by processed for microarray analysis on an Affymetrix Canine Genome 2.0 Array (Affymetrix) using a GeneChip Hybridization, Wash, and Stain Kit (Affymetrix), according to the manufacturer’s instructions. Arrays were scanned by an Affymetrix GeneChip Scanner 3000 System (Affymetrix). The raw data were normalized by Range Migration Algorithm and further analyzed by Affymetrix Microarray Suite Algorithm (MAS5).
Samples RNA was amplified using a GeneChip 3‘IVT Express Kit (Affymetrix, Cleveland, OH, USA) to generate first strand cDNA and second strand cDNA, respectively. Then complementary RNA (cRNA) was produced and labeled using GeneChip HT IVT Labeling Kit (Affymetrix). The labeled cRNA was purified and quality test by an Agilent RNA 6000 Nano Kit (Agilent Technologies), followed by processed for microarray analysis on an Affymetrix Canine Genome 2.0 Array (Affymetrix) using a GeneChip Hybridization, Wash, and Stain Kit (Affymetrix), according to the manufacturer’s instructions. Arrays were scanned by an Affymetrix GeneChip Scanner 3000 System (Affymetrix). The raw data were normalized by Range Migration Algorithm and further analyzed by Affymetrix Microarray Suite Algorithm (MAS5).
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