Info Sheet: Whole Genome Sequencing

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

Workflow

1. QC: DNA quantification using Qubit
2. Library construction from total DNA
3. Library QC
4. Illumina Short-Read Sequencing
5. Bioinformatics data analysis (optional)

Expected turnaround time: 6 weeks after passing QC

Input Requirements

• At least 150 ng high-quality, RNA-free genomic DNA for TruSeq Nano
• At least 20-30 ul volume. We will use 2ul for QC!!
• At least 1'000 ng high-quality (260/280 is 1.8 and 260/230 should be 2.0-2.2),genomic DNA for TruSeq
• Fresh frozen tissue, FFPE, blood samples as source, others on request
• Somatic mutations require control samples from corresponding healthy tissue of the same individual

1.5 ml tubes labeled with B-Fabric Tube ID, normalized to the same concentration

Standard Sequencing Recommendations

NovaSeq; 150bp paired-end reads
Packs of 200M reads
germline mutations: 30-fold coverage
somatic mutations: 70-fold coverage

Supported protocols

PCR-free - highly homogenous genome coverage
TruSeq DNA Nano - standard PCR amplification
Nextera XT - low input, optimized for small genomes, PCR amplicons and plasmids
NEBNext Ultra II - low input, compatible with FFPE DNA samples
Depending on genome size and other needs, long read sequencing may be an alternative

Costs for UZH und ETH research group

Library prep (TruSeq DNA Nano)	108 CHF / sample
Sequencing 200M reads, 300 cycles	1057 CHF
Bioinformatics package	880 CHF

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