MagMAX™-96 Total RNA Isolation Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- This kit can be used for the isolation of RNA from ≤ 5 mg animal tissue.		- To avoid DNA contamination, either reduce sample size, or include an additional DNase treatment step after RNA isolation

Upstream tips
- This kit can be used for the isolation of RNA from ≤ 5 mg animal tissue.
Downstream tips
- To avoid DNA contamination, either reduce sample size, or include an additional DNase treatment step after RNA isolation

Publication protocol

NA from each sample was isolated using MagMAX-96 Total RNA (AM1830; Applied Biosystems), RNA isolation kits, and used as input for library generation with Takara SMARTer Stranded Total RNA Sample Prep Kit (634876; Takara Bio USA). A total of 26 samples were sequenced for 50-bp single-end reads across 10 lanes on an Illumina HiSeq 2500 by the University of Michigan DNA Sequencing Core. All reads were quantified using Kallisto pseudo alignment to an index of transcripts from all human genes within the Ensembl GRCh38 database.12 Gene level data then were generated from the Kallisto estimated counts per transcript using the Bioconductor package tximport.13 Differential expression analysis was performed using the Bioconductor package DESeq2 and the gene level count data. A gene was considered to be differentially expressed if it had a 2-fold or larger difference between groups and an adjusted P value of .01 or less. Principal component analysis and sample clustering were performed in R with log2 transformed and centered gene counts of gene level data on all genes that had a sum of at least 10 counts across all samples. Replicates for all samples were clustered by Euclidian distance, and pairwise Pearson correlation coefficients were plotted in R. All reads are deposited at the EMBL-EBI ArrayExpress archive under accession E-MTAB-6516.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for MagMAX™-96 Total RNA Isolation Kit below.

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