Total RNA from mouse brain tissue was extracted using the Ambion RNAqueous-Micro total RNA isolation kit (Life Technologies Corp.). RNA purity, integrity (RIN > 7) and concentration was determined, and 100 ng of total RNA was then used to synthesize cDNA using SuperScript III (Invitrogen). RT-qPCR was performed on a ViiA7 System (Applied Biosystems) using PerfeCTa SYBR Green (Quanta Biosciences, Gaithersburg, USA). Forward/Reverse primers were designed for different exons, and the RNA was treated with DNase H to avoid false-positives. Amplicon specificity was verified by melting curve analysis. All RT-qPCR reactions were conducted in technical triplicates and the results were averaged for each sample, normalized to Actb levels and the relevant reporter genes such as RFP, GFP and T7 for the viruses, and analyzed using the comparative ΔΔCt method (Livak and Schmittgen, 2001 (link)). The primers (Mus musculus) that were used are listed in Table S3 .
Ambion rnaqueous micro total rna isolation kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Ambion RNAqueous-Micro total RNA isolation kit is a laboratory product designed for the isolation of high-quality total RNA from small samples. It provides a reliable and efficient method for extracting RNA from various sample types, including cells, tissues, and other biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Nextseq 500 platform, by Illumina (2 mentions)
Tg nextseq 500 550 high output kit v2, by Illumina (2 mentions)
Takara smart seq v4 ultra low input rna kit, by Takara Bio (2 mentions)
Viia 7 system, by Thermo Fisher Scientific (1 mentions)
Perfecta sybr green, by Quanta Biosciences (1 mentions)
Superscript 3, by Thermo Fisher Scientific (1 mentions)
Jumpstart taq dna polymerase, by Merck Group (1 mentions)
Affinityscript qpcr cdna synthesis kit, by Agilent Technologies (1 mentions)
4 protocols using ambion rnaqueous micro total rna isolation kit
1
RT-qPCR Analysis of Mouse Brain Transcripts
2
RNA Isolation and cDNA Synthesis for Receptor Analysis
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Total RNA was isolated from macrophages by using the Ambion RNAqueous® Micro total RNA isolation kit (Life Technologies, Carlsbad, CA, USA) in accordance with the instructions of the manufacturer. RNA was reverse-transcribed into complementary deoxyribonucleic acid (cDNA) by using an Affinity Script QPCR cDNA synthesis kit (Agilent Technologies, Santa Clara, CA, USA), and endpoint polymerase chain reaction (PCR) was performed with JumpStart Taq DNA Polymerase (Sigma-Aldrich) and primers for neurotransmitter receptors listed in Additional file 1 : Table S1.
3
Bulk RNA-seq of FAC-sorted Avian Cells
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Total RNAs from FAC-sorted cells were isolated using the Ambion RNAqueous Micro Total RNA isolation kit (Cat #AM1931, ThermoFisher Scientific) and integrity checked using Agilent Tapestation (only samples with a RNA Integrity Number >6 were used for library preparations). Samples were stored at -80ºC until all replicates were collected and sample libraries were prepared on the same day to prevent batch effects. Libraries were prepared using Takara SMART-Seq™ v4 Ultra™ Low Input RNA kit (Cat #634889, Takara Bio Clontech) and sequenced using 40 bp paired-end reads on the Illumina NextSeq 500 platform using TG NextSeq® 500/550 High Output Kit v2 (75 cycles). Two biological replicates were used for each stage (HH10, HH18. HH25) and sample (double positive, single E2-only and Citrine-negative control per stage).
4
Bulk RNA-seq of FAC-sorted Avian Cells
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