Purified total RNA was amplified and labeled using the Low RNA Input Linear Amplification kit (Agilent technologies, Santa Clara, CA, US), 5-(3-aminoallyl)-UTP (Ambion, Austin, TX, US), Cy3 NHS ester (GE healthcare Biosciences, Pittsburgh, PA, US), following the manufacturer’s instructions. Labeled cDNA were purified by RNeasy mini kit (QIAGEN, GmBH, Germany). A designed 8*15K DNA Microarray (Design ID 030218, Agilent), with 8797 probes that covered all the genes of 13 published Salmonella genome, was applied to microarray hybridization. During the hybridization, each slide was hybridized with 1.65 μg of Cy3-labeled cRNA using Gene Expression Hybridization Kit (Agilent technologies, Santa Clara, CA, US) in Hybridization Oven (Agilent technologies, Santa Clara, CA, US), according to the manufacturer’s instructions. After 17 h hybridization, slides were washed in staining dishes (Thermo Shandon, Waltham, MA, US) with Gene Expression Wash Buffer Kit (Agilent technologies, Santa Clara, CA, US), Stabilization and Drying Solution (Agilent technologies, Santa Clara, CA, US), following the manufacturer’s instructions.
5 3 aminoallyl utp
Manufactured by Thermo Fisher Scientific
Sourced in United States
5-(3-aminoallyl)-UTP is a modified nucleotide that can be used in various molecular biology applications. It serves as a precursor for the incorporation of aminoallyl-labeled uridine into RNA molecules.
Automatically generated - may contain errors
Lab products found in correlation
Low rna input linear amplification kit, by Agilent Technologies (2 mentions)
Gene expression hybridization kit, by Agilent Technologies (2 mentions)
Cy3 nhs ester, by GE Healthcare (2 mentions)
Rneasy mini kit, by Qiagen (2 mentions)
Hybridization oven, by Agilent Technologies (2 mentions)
Stabilization and drying solution, by Agilent Technologies (1 mentions)
Gene expression wash buffer kit, by Agilent Technologies (1 mentions)
Cy5 nhs ester, by GE Healthcare (1 mentions)
Feature extraction software 10, by Agilent Technologies (1 mentions)
Gene spring software 11, by Agilent Technologies (1 mentions)
Agilent microarray scanner, by Agilent Technologies (1 mentions)
2 protocols using 5 3 aminoallyl utp
1
Salmonella Genome Microarray Analysis
2
Arabidopsis Microarray Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Microarrays were performed according to a previously described procedure [30 (link)]. Young buds collected from WT and 5mARF17/WT plants were immediately frozen in liquid nitrogen. A Low-RNA-Input Linear Amplification Kit (Agilent Technologies) was used to amplify and label the total RNA. 5-(3-Aminoallyl)-UTP (Ambion), Cy3 NHS ester (GE Healthcare Biosciences) and Cy5 NHS ester (GE Healthcare Biosciences) were applied following the manufacturers’ instructions. The labeled cRNA was purified using an RNeasy Mini Kit (Qiagen). According to the manufacturer’s instructions, each 44-K Arabidopsis oligo microarray slide was hybridized with 825 ng of Cy3-labeled cRNA and 825 ng of Cy5-labeled cRNA using a gene expression hybridization kit (Agilent) in a hybridization oven (Agilent). The slides were scanned using an Agilent Microarray Scanner (Agilent) and Feature Extraction software 10.7 (Agilent) with the default settings. Three biological replicates of independently grown materials were used. The raw data were normalized with a locally weighted scatter plot smoothing (Lowess) algorithm using Gene Spring Software 11.0 (Agilent).
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