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The single-stranded DNA was then fragmented and labelled using the Affymetrix WT Terminal Labelling and Controls Kit, and the distribution of fragment lengths was measured on the Agilent BioAnalyzer.
200 ng RNA was used to generate labelled sense single stranded DNA (ssDNA) for hybridization with the Ambion WT Expression Kit, the Affymetrix WT Terminal Labelling and Controls Kit and the Affymetrix Hybridization, Wash, and Stain Kit as described by the manufacturer.
RNA integrity was assessed on a BioAnalyzer (Agilent Technologies, UK). 200 ng RNA was used to produce labelled sense single stranded DNA (ssDNA) for hybridization with the Ambion WT Expression Kit, the Affymetrix WT Terminal Labelling and Controls Kit and the Affymetrix Hybridization, Wash, and Stain Kit following the manufacturer's instructions.
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Specifically, 4 μg of total RNA was used for reverse transcription and synthesis and amplification of biotin labelled cRNA using the One cycle target labelling and control reagents.
Biotinylated target cRNA was prepared from 50 ng of reverse-transcribed total RNA (One-Cycle Target labelling and control reagents; Agilent Technologies).
Briefly, 100 300 ng of total RNA were amplified and labeled using the WT Sense Target labelling and control reagents kit (Affymetrix Inc., Santa Clara, CA, USA), and then hybridized to GeneChip® Zebrafish Genome Array (Affymetrix).
From each sample >50 ng RNA was extracted using the PicoPure™ RNA isolation kit (Arcturus), and amplified using a 2-cycle linear amplification process, with the GeneChip two-cycle target labelling and control kit (Affymetrix), and MEGAscript® T7 kit (Ambion).
Repetitive high-order digits are removed from the Base Position track, the data are placed front and centre with minimalistic borders and labelling, and control widgets are kept in the margin and in collapsed format until activated by the user.
Its three-year, $10 million grant is being used to support anti-obesity advertising campaigns, finance research at Mexico's National Institute of Public Health and promote policies like the soda tax, nutrition labeling and controls on junk-food television advertising aimed at children.
Sense ssDNA was fragmented and labeled using the GeneChip WT Terminal Labeling and Controls Kit.
Target preparation, including hybridization and post-hybridization procedures, was performed as described by the Affymetrix GeneChip Whole-transcriptome Terminal Labeling and Controls manual (http://www.affymetrix.com).
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