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In case of fat tissues, the RNeasy lipid kit (Qiagen) was applied.
Total RNA was extracted from 250 μg tissue samples by homogenization in QIAzolLysis Reagent (Qiagen) and according to RNeasy Lipid kit (Qiagen) protocol.
We used RNeasy kit or RNeasy lipid kit (fat-rich tissues) for total RNA extraction according to the manufacturer's instructions (Qiagen).
Total RNA was isolated from adipose tissue using the Trizol RNA isolation method (Invitrogen, Carlsbad, CA) and purified with the RNeasy Lipid kit (QIAGEN -Ref. 74804-, ValenCAa, CA).
Total RNA was isolated from chicken adipose samples using the RNeasy Lipid kit and incorporating an on-column DNase treated with the RNase-free DNase Set according to the manufacturer's protocol (Qiagen.com).
RNA was isolated (QIAGEN RNeasy Lipid Kit, Australia) from Cape fur seal, wallaby, pig and mouse mammary tissue or from cells derived from California sea lion and Antarctic fur seal milk during peak lactation, human milk and colostrum at 24 hours prior to birth and at 3 and 4 months peak lactation, or HEK293T cells.
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Total RNA was isolated using two different commercial kits: RNeasy Lipid Tissue Kit (QIAGEN) for the fat-rich cerebrum and RNeasy Mini Kit (QIAGEN) for the liver.
RNA isolation was performed using RNeasy Lipid Tissue kit and RNeasy Mini kit (Qiagen, Valencia, CA, USA) according to the manufacturer's protocols.
Different RNA extraction kits and protocols were used for processing different tissues: QIAGEN RNeasy® Lipid Tissue Kit for cerebral cortex samples, QIAGEN RNeasy® Kit for liver samples, and QIAGEN RNeasy® Fibrous Tissue Kit for skeletal muscle samples.
Real-Time Quantitative PCR Embryonic (E10.5, E12.5, E14.5, E16.5, E18.5), postnatal (P0, P6) and adult (8 weeks) tissues (C57/BL6) were dissected and snap frozen, before mRNA extraction employing either an RNeasy lipid tissue kit or an RNeasy fibrous tissue kit.
Total RNA was extracted using the RNeasy Mini Lipid extraction kit (Qiagen) and 5 μl was reverse transcribed for 1 hour at 45°C using the AffinityScript kit (Agilent) in a 25 μl reaction.
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