Total RNA from heart tissues was extracted using an Aurum Fatty and Fibrous Tissue kit (Biorad, 732–6870), and RNA from NRVMs was isolated using a Quick-RNA MicroPrep kit (Zymo Research, R1051). Reverse transcription was performed using iScript Reverse Transcription Supermix (Biorad) according to the manufacturer’s instructions. mRNA levels were determined by quantitative PCR using a LightCycler machine (Roche) and the iTaq Universal SYBR Green Supermix (Biorad). All primer sequences are listed in Table I in the Supplement .
Aurum fatty and fibrous tissue kit
Manufactured by Bio-Rad
The Aurum Fatty and Fibrous Tissue Kit is a laboratory tool designed for the extraction and purification of nucleic acids from fatty and fibrous tissue samples. The kit includes reagents and components necessary for the isolation process.
Automatically generated - may contain errors
Lab products found in correlation
Iscript reverse transcription supermix, by Bio-Rad (2 mentions)
Itaq universal sybr green supermix, by Bio-Rad (2 mentions)
Quick rna microprep kit, by Zymo Research (2 mentions)
Lightcycler machine, by Roche (2 mentions)
Truseq reagents, by Illumina (1 mentions)
Bioanalyzer, by Illumina (1 mentions)
3 protocols using aurum fatty and fibrous tissue kit
1
Quantitative RNA Analysis of Heart Tissues
2
Quantifying Cardiac Gene Expression
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Total RNA from heart tissues was extracted using an Aurum Fatty and Fibrous Tissue Kit (732-6870; Bio-Rad) and RNA from NRVMs was isolated using a Quick-RNA MicroPrep Kit (R1051; Zymo Research). Reverse transcription was performed using iScript Reverse Transcription Supermix (Bio-Rad) according to the manufacturer’s instructions. mRNA levels were determined by quantitative PCR using a LightCycler machine (Roche) and the iTaq Universal SYBR Green Supermix (Bio-Rad). All primer sequences are listed in Table I in the Data Supplement .
3
Transcriptome Profiling of Copepod Development
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Total RNA was extracted from copepods using the Aurum Fatty and Fibrous Tissue Kit (Bio-Rad). RNA purity and yield were assessed using a Nanodrop spectrophotometer. For the time series study, on-column DNase digestion was used to remove genomic DNA, and RNA quality was assessed using denaturing agarose gels. For the Illumina study, RNA quality was assessed using a Bioanalyzer. Samples were submitted to the Genomic Services Laboratory at HudsonAlpha (Huntsville, AL), where libraries were synthesized using Illumina TruSeq reagents. Four libraries were constructed from each of days 3 and 10, and each library was prepared using total RNA extracted and pooled from enough copepods to yield at least 4 μg of RNA (3–6 copepods). For transcriptome assembly, an additional library was constructed from equal amounts of RNA pooled from each of these eight samples (2 μg total) along with 2 μg of RNA from wild C5 C. finmarchicus collected in Trondheimsfjord, Norway during May 2012. RNA from wild copepods was included in the library to facilitate future field-based transcriptional profiling studies.
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