DNA Library Preparation and Sequencing
Our DNA services includes whole genome, whole exome or targeted library preparation and sequencing on Illumina platforms. See below for available workflows.
We also provide consultation services to lend our experience to researchers and we are open to collaboration on protocols currently under development.
Human Whole Genome Sequencing
- Illumina TruSeq DNA PCR-Free Library Prep Kit as standard workflow
- Illumina TruSeq DNA Nano Library Prep Kit for low input samples
Whole Exome Sequencing and Targeted DNA Sequencing
Libraries are prepared using:
- Roche KAPA HyperPlus Kit with UMI-containing adapters
Enrichment is performed using:
- Twist Comprehensive Exome Probe Panel for Whole Exome Sequencing
- Any Twist Custom Probe Panel or predefined probe panel targeting genomic area of interest
Small Genome Sequencing
- Illumina DNA Prep, Tagmentation Kit (previously called Nextera DNA)
If you are interested in collaborating with us on development of new protocols, or if you have a suggestion for the implementation of a novel method, please contact tina [dot] catela_ivkovic [at] med [dot] lu [dot] se (Tina Catela Ivkovic).
General information before starting a DNA Sequencing project
- DNA should be of high quality and diluted in water or EB buffer, presence of EDTA in the storage buffer might affect the library preparation.
- DNA isolated from FFPE samples must be of high quality, QC needs to be performed by a qPCR-based method (available at CTG).
- DNA should be of high purity, recommended OD260/280 of 1.8-2.0 and OD260/230 of 1.8-2.2 based on spectrophotometric measurement (e.g. by Nanodrop).
- All samples must be diluted/normalized based on concentration measurements performed by a fluorometric method (e.g. Qubit). Measurements based on spectrophotometric method (e.g. Nanodrop) are not acceptable.
- When requesting a project in iLab please specify the required sequencing depth for each sample.
For Whole Exome and Targeted DNA Sequencing
- Up to eight samples/libraries are pooled prior to the enrichment step.
- The price for this particular part of the project is calculated per pool. (For example, if you have 9 samples this would be calculated as two pools).
- If you are interested in custom probe pools for targeted sequencing, contact us for more information.
- DNA input amounts differ between assays. Please see the table below for required sample concentration and delivery volumes. Lower concentrations and volumes can be discussed.
- Always dilute samples in nuclease-free H2O or EB buffer.
|TruSeq DNA PCR-Free||25 ng/µl||100 µl||2.5 µg|
|TruSeq DNA Nano||2 ng/µl||100 µl||200 ng|
|KAPA-Twist||10 ng/µl||30 µl||300 ng|
|Illumina DNA Prep||10 ng/µl||30 µl||300 ng|
Delivery of samples to CTG
DNA should be delivered aliquoted on ice in:
- an 8-tube strip labeled with Sample ID or Sample number (i.e. 1, 2, 3...) on the lid and side of the tube.
- a fully skirted 0.2 ml plate with cold resistant seal (can be provided by CTG). Samples bust be placed column-wise (A1, B1, C1...)
Requesting a project at CTG
To initiate a project using our available services, please start a project in iLab. You can login using your LucatID (SWAMID) on the Lund University iLab webpage.
Data management and additional analysis
CTG uses secure LSENS servers hosted by LUNARC, center for scientific and technical computing at Lund University facility, for internal data management and analysis.
- All data is by default delivered to the customer via a dedicated secure file server hosted on Lund University.
- For an additional cost, data can be delivered on a hard disk.
- For alternative options, please send us an inquiry.
After data delivery, data will be kept by CTG for 3 months and subsequently deleted. We strongly recommend you to backup your data, i.e. have at least two copies. CTG cannot help you if your data gets corrupt or lost!
For additional Bioinformatics services please contact NBIS – National Bioinformatics Infrastructure Sweden, (https://nbis.se/).
Contact for service
E-mail: CTGservice [at] med [dot] lu [dot] se (CTGservice[at]med[dot]lu[dot]se)