Info Sheet: Whole Genome Sequencing
How to submit samples for Illumina/Short Read Sequencing.pdf
Workflow
- QC: DNA quantification using Qubit
- Library construction from total DNA
- Library QC
- Illumina Short-Read Sequencing
- Bioinformatics data analysis (optional)
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
Standard Sequencing Recommendations
NovaSeq; 150bp paired-end readsPacks of 200M reads
germline mutations: 30-fold coverage
somatic mutations: 70-fold coverage
Supported protocols
PCR-free - highly homogenous genome coverageTruSeq 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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