RNA Profiling

RNA Sequencing using Illumina Nextseq500 platform

 

Sample requirement:

  • cDNA: Submit DNase treated RNA of approximately 200-500ng of high quality with Nanodrop A260/280 ratio >1.9; A260/230=2.0-2.2; and at a concentration of ~20 ng/ul (dissolve in nuclease free water).Supplied RNA should be shipped in dry ice to the Eurofins facility.
  • cDNA should be more than 80% of provided RNA.

 

Sample Quality Control: 

The qualification and quantification of cDNA will be carried out by denatured agarose gel electrophoresis and NanoDrop/Qubit Fluorometer analysis.

 

Library Preparation: 

  • RNA Seq library will be prepared by using TruSeq Stranded mRNA Sample Prep Kit or TruSeq Stranded Total RNA Library Prep Kit.
  • Preparation of indexed RNA library by enriching mRNA from Total RNA for sequencing on Illumina Platform.
  • Library validation will be carried out using High Sensitivity D1000 Screen tape using Agilent Tape Station.

 

Sequencing: 

  • Sequencing will be carried out on Illumina Nextseq500 platform with 2×150 for RNA v2 chemistry. Generation of data per sample.

 

Bioinformatics Analysis:

  • Quality filtration of reads,
  • Mapping on the reference genome ( as provided by customer),
  • Alignment summary statistics,
  • Gene annotation
  • Compiled Report

 

Delivery Time :

Provided that there are no unexpected difficulties (biological or technical), 5-6  weeks from the date of QC qualified samples received. All samples to be provided in a single lot.

 

Confidentiality :

Eurofins Genomics will perform the services outlined above on a “fee for service” basis. This means that Eurofins Genomics has and will claim no rights whatsoever to all or any of the data that will be generated. Eurofins Genomics will take all necessary measures to ensure that none of the data will be disseminated to third party.