Single strand cRNA was labeled with Cyanine 5-CTP or Cyanine 3-CTP (Agilent, USA). Reaction efficiency and activity were determined by NanoDrop. Hybridization to the Bovine (V2) Gene Expression Microarrays 4 × 44 K (Agilent, USA), washing, and scanning was performed according to the Two-Color Microarray-Based Gene Expression Analysis (Quick Amp Labeling) with Tecan HS Pro Hybridization. Following 17 h of hybridization, arrays were scanned on an Agilent G2565AA scanner. Images were quantified using Agilent Feature Extraction Software (version A.8.5.1.1). Pooling, labeling, and hybridization of RNA sample schemes in the microarray analysis are shown in Additional file 12 .
Cyanine 3 ctp
Manufactured by Agilent Technologies
Sourced in United States
Cyanine 3-CTP is a fluorescent labeling reagent used in various molecular biology applications. It is a cyanine dye that can be incorporated into nucleic acid molecules, enabling their detection and visualization. The core function of Cyanine 3-CTP is to provide a fluorescent label for nucleic acid samples.
Automatically generated - may contain errors
Lab products found in correlation
Feature extraction software, by Agilent Technologies (3 mentions)
Bovine v2 gene expression microarrays 4 44 k, by Agilent Technologies (1 mentions)
G2565aa scanner, by Agilent Technologies (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Genespring, by Agilent Technologies (1 mentions)
Agilent scanner g2505c, by Agilent Technologies (1 mentions)
3 protocols using cyanine 3 ctp
1
Bovine Transcriptome Analysis by Microarray
2
Comparative lncRNA profiling of osteoclasts
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Undifferentiated RAW264.7 cells and RAW264.7-derived osteoclasts on day 4 were lysed using TRIzol Reagent (Invitrogen), frozen in dry ice, and sent to Oebiotech (Shanghai, China) for lncRNA microarray analysis. Briefly, total RNA was transcribed to double-stranded cDNA, synthesized into cRNA and labelled with cyanine-3-CTP (Agilent Technologies, Santa Clara, USA). The labelled cRNAs were hybridized onto the Agilent mouse lncRNA microarray (4*180K; Agilent Technologies). After washing, the arrays were scanned with an Agilent Scanner G2505C (Agilent Technologies). Feature Extraction software (version 10.7.1.1; Agilent Technologies) was used to analyze array images to obtain raw data. Genespring (version 13.1; Agilent Technologies) was employed to complete the basic analysis with the raw data. The threshold set for up- and downregulated genes was a fold change ≥ 2.0 and a
P value ≤ 0.05.
P value ≤ 0.05.
3
Colorectal Tissue Transcriptome Analysis
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An analysis of 55 RNA samples derived from colorectal tissue, consisting of three sample groups (NOR, CRA and CRC) with varying numbers of biological replicates, was performed on 44k Whole Human Genome microarrays (Agilent) that contain 41,093 probes, providing full coverage of human transcripts. Double-stranded cDNA was synthesized from 500 ng of total RNA using the Quick Amp Labeling kit, One-color, as instructed by the manufacturer (Agilent). Labeling with cyanine3-CTP, fragmentation of cRNA, hybridization, and washing were performed according to the manufacturer’s instructions (Agilent). The microarrays were scanned and the data were extracted with the Agilent Feature Extraction Software.
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