Total RNA was obtained after 9 h of fermentation using a Total RNA Isolation Mini Kit (Agilent Technologies) according to the manufacturer’s protocol. The concentration and quality of RNA were determined using a NanoDrop ND-1000 spectrophotometer (NanoDrop, Wilmington, DE) and an Agilent 2100 Bioanalyzer (Agilent Technologies), respectively. cDNA was reverse transcribed and labeled with cyanine 3-CTP using a Low-Input Quick Amp Labeling Kit (Agilent Technologies) for hybridization to S. cerevisiae 4 × 44 k microarrays (Agilent Technologies). Hybridization was performed at 65 °C for 17 h. The arrays were scanned using an Agilent Single-Color DNA Microarray Scanner (Agilent Technologies). Gene expression levels were normalized per chip. Scan data were analyzed using GeneSpring GX ver. 11.5.1 software (Agilent Technologies). The experiment was performed in triplicate. The accession number of the microarray data was GSE73814.
Agilent single color dna microarray scanner
Manufactured by Agilent Technologies
The Agilent Single-Color DNA Microarray Scanner is a laboratory instrument designed to detect and quantify fluorescent signals from DNA microarray samples. It is capable of scanning single-color microarray slides and producing high-resolution digital images of the microarray data.
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2 protocols using agilent single color dna microarray scanner
1
Transcriptome Analysis of S. cerevisiae
2
Transcriptome analysis of microbial fermentation
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Total RNA was obtained after 3, 6, 24, and 48 h of fermentation using a Total RNA Isolation Mini Kit (Agilent Technologies) according to the manufacturer's protocol. RNA concentration and quality were measured using a NanoDrop ND-1000 spectrophotometer (NanoDrop, Wilmington, DE) and an Agilent 2100 Bioanalyzer (Agilent Technologies), respectively. cDNA was reverse-transcribed and labeled with cyanine 3-CTP using a Low-Input Quick Amp Labeling Kit (Agilent Technologies) for hybridization in 8×15k microarrays designed by the author. Hybridization was performed at 65 °C for 17 h and the arrays were scanned using an Agilent Single-Color DNA Microarray Scanner (Agilent Technologies). Gene expression levels were normalized per chip. GeneSpring GX ver. 11.5.1 software (Agilent Technologies) was used to analyze the fold-change in expression data. Each biological sample subjected to microarray analysis was analyzed in triplicate.
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