Trizol reagent protocol
TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other components designed for the isolation of total RNA from various biological samples. The reagent maintains the integrity of RNA while disrupting cells and dissolving cell components.
Lab products found in correlation
79 protocols using trizol reagent protocol
Quantitative RT-PCR for Lung RNA Analysis
Tambaqui Liver RNA Extraction and Reverse Transcription
the TRIzol® reagent protocol (Invitrogen) according to the manufacturer’s
instructions. Contaminating genomic DNA was removed using DNase I (Invitrogen).
First strand cDNA was reverse-transcribed from the total RNA using RevertAid H Minus
First Strand cDNA Synthesis kit (Fermentas), and following the manufacturer’s
instructions. Enzymatic treatment with reverse transcriptase (MMLV Reverse
Transcriptase) (200 U/μL, USB) was first done, and then mixed in a 0.2 mL microtube
with approximately 25 μg RNA, 1 μL of Oligo(dT)18 primer (1 μg), 1.0 μL
dNTP mix (10 mM), MMLV buffer 5X, and deionized water for a 20 μL final volume. This
solution was incubated at 37 °C for 1 h for conversion and at 70 °C for 10 min to
inactivate the enzyme.
Embryonic RNA-Seq Library Preparation
Sample quality was assessed using a 2100 Bioanalyzer 2100 (Agilent Technologies) (RIN > 8.5). Library preparation was performed using the HTP RNA-Seq Library Prep Kit (iGenomX Inc, San Francisco, CA, USA). Briefly, barcoded oligo dT primers were added to 50 ng RNA per sample and reverse transcribed. Sample cDNA products were combined, cleaned, and run through PCR with 0.5 µM barcoded PCR primers (p5 and p7 sequences, Illumina, San Diego, CA, USA). Purified PCR products were sequenced using a NextSeq2000 sequencer (paired-end mode; read1: 26 bp, read2: 94 bp), generating greater than 1.5 million paired-end reads for each sample.
Transcriptomic Analysis of Soybean-Sclerotinia Interaction
Library preparation was performed at the University of Wisconsin – Madison Biotechnology Centre (Madison, WI, USA). Individually indexed libraries were prepared using the TruSeq RNA Sample Preparation v2 kit according to the manufacturer’s instructions (Illumina, San Diego CA, USA). Library concentrations were quantified with the Qubit HS DNA kit (Thermo Fisher Scientific, Wilmington, DE). The size and quality of the libraries were evaluated with an Agilent Bioanalyzer 2100 and an Agilent DNA 1000 kit (Agilent Technologies, Santa Clara, CA) and the libraries were sequenced using Illumina HiSeq2500 (1X100bp) (Illumina, San Diego CA, USA).
Quantitative Gene Expression Analysis
Rumen Microbiome RNA Isolation and Sequencing
Kidney mRNA Expression Quantification
Quantifying Drosophila Transcript Levels
Quantifying FRα mRNA Expression in Cancer
RNA Extraction and cDNA Synthesis Protocol
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