Endocannabinoid System Microarray Analysis

For the microarray analysis, 500 ng of total RNA was converted to double stranded cDNA and this was used to generate biotinylated cRNA probes using the Illumina TotalPrep RNA Amplification Kit. Biotin-labelled cRNA were then hybridized to Illumina RatRef-12 Expression BeadChip (San Diego, CA, USA). Slides were scanned on a BeadStation 500 System using Beadscan software Version 3.5.31. Each of the placental samples was analysed independently. Array experiment readout was deposited on ArrayExpress (ArrayExpress accession number E-MTAB-1987). Overall microarray results were previously published by our group [16 (link)]. For this study, expression of the 7 enzymes Faah, Mgll, Plcd4, Pld1, Nat1, Daglα, and Ptgs2 in the endocannabinoid system was analyzed.

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Vaswani K., Chan H.W., Peiris H.N., Dekker Nitert M., Wood Bradley R.J., Armitage J.A., Rice G.E, & Mitchell M.D. (2015). Gestation Related Gene Expression of the Endocannabinoid Pathway in Rat Placenta. Mediators of Inflammation, 2015, 850471.

Publication 2015

TotalPrep RNA Amplification Kit RatRef-12 Expression BeadChip

Biotin Cdna Crna Endocannabinoid Enzymes Microarray Nat1 Placental Pld1 Ptgs2

Corresponding Organization : University of Queensland

Other organizations : Monash University, Deakin University

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Variable analysis

independent variables

Amount of total RNA (500 ng)

dependent variables

Expression of the 7 enzymes Faah, Mgll, Plcd4, Pld1, Nat1, Daglα, and Ptgs2 in the endocannabinoid system

control variables

Use of Illumina TotalPrep RNA Amplification Kit to convert total RNA to double stranded cDNA and generate biotinylated cRNA probes
Hybridization of biotin-labelled cRNA to Illumina RatRef-12 Expression BeadChip
Scanning of slides on a BeadStation 500 System using Beadscan software Version 3.5.31
Independent analysis of each placental sample

controls

No positive or negative controls were explicitly mentioned in the provided information.

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