Info Sheet: Small RNA Sequencing

How to submit samples for Illumina/Short Read Sequencing.pdf

Workflow

  1. QC: RNA quantification and checking the RNA integrity
  2. Library construction from total RNA using the Illumina TruSeq Small RNA Library Kit
  3. Library QC
  4. Sequencing
  5. Bioinformatics data analysis (optional)
Expected turnaround time: 6-8 weeks after passing QC

Input requirements

  • at least 200 ng/ul high-quality total RNA, inquire for low input protocols
  • RNA must be purified and DNA-free. Recommended RNA extraction protocols:
    • Trizol purification
    • Qiagen miRNeasy (or similar)
1.5 ml tubes labeled with B-Fabric Tube ID, normalized to the same concentration

Experimental Design

3 or more replicates for cell lines, well-defined tissues or organs in model organisms
5 or more replicates for human samples, low input samples

Standard Sequencing Recommendations

75 bp single-end reads, 10M reads / sample
Packs of 200M reads

Bioinformatics

Read alignment, differential analysis, functional analysis and visualization available via https://fgcz-sushi.uzh.ch/
Additional analyses available as custom scripts

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