FGCZ Genomics > Services> NGS Services

Info Sheet: total RNA-seq


Workflow

  1. QC: RNA quantification and checking the RNA integrity
  2. Library construction from total RNA using ribo-depletion
  3. Library QC
  4. Sequencing
  5. Bioinformatics data analysis (optional)
Expected turnaround time: 6 weeks after passing QC

Input requirements

  • at least 300 ng total RNA + 3 ul for QC
  • at least 15 μl sample volume
  • concentration between 30 ng/μl - 100 ng/μl
  • RNA must be DNA free (e.g. DNase treated)
  • RNA from whole blood have special requirements (we must deplete for globin)!
  • bacterial RNA should first be depleted of ribosomal RNA
  • RNA should have RIN > 2, inquire for highly degraded RNA
1.5 ml tubes labeled with B-Fabric Tube ID

Experimental design

3 or more replicates per condition (e.g. cell lines, mouse models, …)
5 or more replicates per condition (e.g. humans, high variability, low quality RNA, elaborate/complex extraction)

Standard Sequencing Recommendations

NovaSeq; 100bp single-end reads; 40M reads/sample
Packs of 500M reads

Costs for UZH and ETHZ research groups

Library preparation 190 CHF / sample
Sequencing 500M reads (up to 12 samples) 788.80 CHF
Bioinformatics (up to 15 samples, 3 comparisons) 880 CHF


Ordering

Instructions can be found here

Alternatives
mRNA-seq using poly-A selection
single-cell RNA-seq
ribosome profiling
higher sequencing depth
low input protocols (1 ng - 10 ng)


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