10 primiparous lactating Holstein cows (body weight, 558 ± 10 kg; days in milk, 136 ± 37d; daily milk yield, 21.1 ± 2.3 kg) were fed with 40% forage and 60% concentrate (CS; n=5) or 60% forage and 40% concentrate (MF; n=5) for 9 weeks. At the end of 9 weeks biopsy from mammary and liver were obtained.
Extracted molecule
total RNA
Extraction protocol
RNA was extracted with TRIZOL Reagent (Cat#15596-018, Life technologies, Carlsbad, CA, US). Total RNA was purified by Rneasy mini kit (QIAGEN, Germany, Cat#74106) and RNase-Free DNase Set (QIAGEN, Germany, Cat#79254). The quality of RNA was analyzed with Agilent Bioanalyzer 2100 (Agilent technologies, Santa Clara, CA, US).
Label
Cy3
Label protocol
Total RNA was amplified and labeled by Low Input Quick Amp Labeling Kit, One-Color (Agilent technologies, US Cat#5190-2305) according to the manufacturer’s instructions.
Hybridization protocol
Each slide was hybridized with 1.65 μg Cy3-labeled cRNA using Gene Expression Hybridization Kit (Agilent technologies, US, Cat#5188-5242). Hybridization was carried out with a hybridization oven (Agilent technologies, US) for 17 hours.
Scan protocol
Slides were scanned by Agilent Microarray Scanner (Agilent technologies, US) with default settings (i.e., dye channel: Green, Scan resolution= 5 μm, PMT, 100%, 10%, 16bit).
Description
Gene expression of liver sample
Data processing
Data was acquired with Feature Extraction software 10.7 (Agilent technologies, US) and normalized by Quantile method with Gene Spring Software 11.0 (Agilent technologies, US)
