High-quality genomic coverage from nasopharyngeal, oropharyngeal, and diverse samples increases accuracy in SARS-CoV-2 research such as the study of viral evolution and transmission patterns. The optimized workflow for SARS-CoV-2 reduces time and sequencing costs while providing flexibility to use available reagents.
Improve library generation for high-throughput sequencing with high yields and strong coverage of biologically important genes with the Invitrogen Collibri PCR-Free Enzymatically Sheared (ES) DNA Library Preparation Kit for Illumina Systems or the Invitrogen Collibri ES DNA Library Preparation Kit for Illumina Systems.
All components, including shearing enzymes, indexed adaptors, amplification master mix (if relevant), and cleanup beads are included for convenience and performance. Unique Dual Indexes (UDIs) are available for 24 or 96 reactions to filter index-hopped reads on patterned flow cells.
Sequence SARS-CoV-2 samples
PCR is a choice
Yield from PCR-free Collibri ES DNA protocols is optimized to convert 100 ng of DNA into sufficient amount of library to perform quantification and more than two runs on an Illumina™ NovaSeq™ system. Resulting libraries contain fewer chimeric reads than older library prep technology.
Figure 1. Coriell NA12878 DNA was converted into sequencing libraries using the manufacturer’s recommended PCR-free protocols and concentration was measured by the Collibri Library Quantification kit. Libraries were sequenced on an Illumina NovaSeq 6000 System.
Strong coverage of biologically important genes
Consistent gene coverage of biologically important genes enables greater discovery power without additional sequencing depth. Improve sensitivity of variant detection from low inputs of DNA compared to older library prep technology.
Click image to enlarge
Figure 2. 1 ng of Coriell NA12878 DNA was converted into a library following manufacturer instructions, sequenced on a 2x150 bp paired end run on an Illumina NovaSeq 6000, and reads normalized to 258 m per sample. Coverage was calculated by Picard Tools and SNPs were detected by Freebayes v1.2.0.
Whole-genome library preparation workflow
Genomic DNA extraction
We offer a range of Invitrogen genomic DNA extraction kits for sensitive, scalable purification from an expansive set of starting materials to help maximize process efficiency and downstream performance. This includes a broad range of kits for purifying genomic DNA from a variety of samples including tissue, cells, blood, serum, plants, forensic samples, and more.
Automate DNA extraction using magnetic bead-based technology and KingFisher instruments to bind, wash, and elute DNA to reduce your hands-on time while maintaining high yields and excellent reproducibility.
Sensitive and specific nucleic acid quantification
The Qubit dsDNA high-sensitivity (HS) and broad-range (BR) assays are highly selective for double-stranded DNA (dsDNA) over RNA and single-stranded DNA (ssDNA). The high-sensitivity assay is recommended for samples with a low concentration of dsDNA, making it ideal for precious samples.
Recommended accessories
- Interchange the 96-well 0.2-mL block with another block type if your throughput needs change
- Access the system remotely (and conveniently) through a mobile app
- Program the instrument in seconds with a simple-to-use touchscreen interface
- VeriFlex blocks allow for precise PCR optimization or to run up to six independent assays
- Easy-to-use graphical interface (6.5-inch VGA Touch Screen)
- Reduced PCR cycling time when using faster ramp rates
Quantification options to scale throughput
The qPCR-based Collibri Library Quantification kit scales well for larger sample batches and is the ideal method for precious samples or clinical samples. Quantification accuracy is equivalent to the KAPA Library Quantification kit with the additional benefit of visual feedback during the quantification process.
The Qubit dsDNA HS Assay is a fluorometric assay that uses dsDNA-binding dyes in order to accurately determine NGS library concentration, and benefits from a simple workflow of just a few minutes per sample.
Regardless of the assay that is chosen, good laboratory technique should be used in order to ensure accurate measurement of library concentrations and high-quality Illumina sequencing data.
Recommended accessories
With Applied Biosystems QuantStudio real-time PCR systems, you get true value with excellent performance, reliability, and world-class support. Our family of instruments enables you to obtain the results you need, connect and collaborate with colleagues, and achieve your research goals.
Genomic DNA extraction
We offer a range of Invitrogen genomic DNA extraction kits for sensitive, scalable purification from an expansive set of starting materials to help maximize process efficiency and downstream performance. This includes a broad range of kits for purifying genomic DNA from a variety of samples including tissue, cells, blood, serum, plants, forensic samples, and more.
Automate DNA extraction using magnetic bead-based technology and KingFisher instruments to bind, wash, and elute DNA to reduce your hands-on time while maintaining high yields and excellent reproducibility.
Sensitive and specific nucleic acid quantification
The Qubit dsDNA high-sensitivity (HS) and broad-range (BR) assays are highly selective for double-stranded DNA (dsDNA) over RNA and single-stranded DNA (ssDNA). The high-sensitivity assay is recommended for samples with a low concentration of dsDNA, making it ideal for precious samples.
Recommended accessories
- Interchange the 96-well 0.2-mL block with another block type if your throughput needs change
- Access the system remotely (and conveniently) through a mobile app
- Program the instrument in seconds with a simple-to-use touchscreen interface
- VeriFlex blocks allow for precise PCR optimization or to run up to six independent assays
- Easy-to-use graphical interface (6.5-inch VGA Touch Screen)
- Reduced PCR cycling time when using faster ramp rates
Quantification options to scale throughput
The qPCR-based Collibri Library Quantification kit scales well for larger sample batches and is the ideal method for precious samples or clinical samples. Quantification accuracy is equivalent to the KAPA Library Quantification kit with the additional benefit of visual feedback during the quantification process.
The Qubit dsDNA HS Assay is a fluorometric assay that uses dsDNA-binding dyes in order to accurately determine NGS library concentration, and benefits from a simple workflow of just a few minutes per sample.
Regardless of the assay that is chosen, good laboratory technique should be used in order to ensure accurate measurement of library concentrations and high-quality Illumina sequencing data.
Recommended accessories
With Applied Biosystems QuantStudio real-time PCR systems, you get true value with excellent performance, reliability, and world-class support. Our family of instruments enables you to obtain the results you need, connect and collaborate with colleagues, and achieve your research goals.
Ordering information
Resources
App note
Brochures
DNA sequencing
Technical notes
All sequencing methods
Technical notes
RNA sequencing
Technical notes
For Research Use Only. Not for use in diagnostic procedures.