TruSeq RNA Library Prep Kit v2

RNA sequencing Human - Huh7

Experiment
RNA sequencing Human - Huh7
Product
TruSeq RNA Library Prep Kit v2 from Illumina
Manufacturer
Illumina

Protocol tips

Protocol tips
0.1 - 1 ug total RNA or 10 - 400 ng previously isolated mRNA (from species with polyA tails)

Publication protocol

TCR-redirected or mock-treated T cells cocultured with Huh7A2 HCVRep cells at an E/T ratio of 10:1 were collected at the indicated times and enriched with anti-CD8 MicroBeads (Miltenyi Biotech) as per the manufacturer's instructions. Enriched CD8+ T cells (purity, 98% CD3+ CD8+) at 1.5 million cells per sample were stored at −80°C. Total RNA was isolated with TRIzol reagent (Thermo Fisher Scientific, MA, USA), and RNase-free reagents were used throughout the isolation. Sample quantity and purity were assessed using the NanoDrop 1000 spectrophotometer (Thermo Fisher Scientific, MA, USA). All RNA samples had RNA integrity number (RIN) values of >9. The RNA libraries were bar coded and prepared according to the protocol Illumina TruSeq RNA Sample Prep kit v2 (catalog number RS-122-2001) from the manufacturer (Illumina, San Diego, CA, USA).
The nine RNA libraries were sequenced in paired ends on an Illumina HiSeq 2000 instrument, where each sample was put in two different lanes, for a total of 18 samples. The runs generated a total of 384 million reads with an average of 43 million paired-end reads per sample, of which 90% mapped to the human genome. The paired-end sequences were aligned to the human genome reference hg19, with TopHat2 version 0.9 using TopHat standard parameters. The numbers of aligned reads per gene were determined with HTSeq 0.6.1. Annotations from Ensembl Homo sapiens grch37.82 and Genome Browser were used to assign features to genomic positions. The R/Bioconductor package DESeq2 was used for differential gene expression on read counts generated by HTSeq 0.6.1.

Full paper   Login or join for free to view the full paper.

Reviews

TruSeq RNA Library Prep Kit v2 from Illumina has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA sequencing Human - Huh7 using TruSeq RNA Library Prep Kit v2 from Illumina.

Paper title
Hepatitis C Virus-Specific T Cell Receptor mRNA-Engineered Human T Cells: Impact of Antigen Specificity on Functional Properties
Full paper
Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Illumina for TruSeq RNA Library Prep Kit v2 below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing RNA sequencing Human - Huh7 using TruSeq RNA Library Prep Kit v2 from Illumina. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Become shareholder Discussions About us Contact Privacy Terms