Murine organoids were grown for 6 days in media containing 1 nM DHT and then for 24 hr without DHT and EGF. Next, cells were stimulated with 10 nM DHT without EGF for 24 hr and harvested for RNA-seq in biological quadruplicates. Total RNA was extracted, and DNaseI treated by Maxwell 16 LEV simplyRNA Cells Kit and Maxwell nucleic-acid extraction instrument (Promega). Nanodrop quantified RNA was checked by Bioanalyzer RNA 6000 Nano Kit (Agilent Technologies). Samples with RNA integrity number > 10 were used for library preparation (TruSeq Stranded mRNA Library Preparation for Poly-A selection and Stranded RNA-Seq) at the WCM Genomics Core. Libraries were sequenced twice on NextSeq500 (Illumina), High-output mode to generate 75 bp reads at WCM Genomics Core.
Maxwell nucleic acid extraction instrument
Manufactured by Promega
The Maxwell nucleic-acid extraction instrument is a versatile automated system designed to extract high-quality nucleic acids from a variety of sample types. The instrument utilizes magnetic bead technology to efficiently purify DNA, RNA, or other nucleic acids, providing a consistent and reliable extraction process.
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2 protocols using maxwell nucleic acid extraction instrument
1
Murine Organoid RNA-seq Analysis
2
Murine Organoid RNA-seq Analysis
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Murine organoids were grown for 6 days in media containing 1 nM DHT and then for 24 hr without DHT and EGF. Next, cells were stimulated with 10 nM DHT without EGF for 24 hr and harvested for RNA-seq in biological quadruplicates. Total RNA was extracted, and DNaseI treated by Maxwell 16 LEV simplyRNA Cells Kit and Maxwell nucleic-acid extraction instrument (Promega). Nanodrop quantified RNA was checked by Bioanalyzer RNA 6000 Nano Kit (Agilent Technologies). Samples with RNA integrity number > 10 were used for library preparation (TruSeq Stranded mRNA Library Preparation for Poly-A selection and Stranded RNA-Seq) at the WCM Genomics Core. Libraries were sequenced twice on NextSeq500 (Illumina), High-output mode to generate 75 bp reads at WCM Genomics Core.
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