Figure 1. Coriell NA12878 DNA was converted into a library and sequenced on an Illumina™ NovaSeq™ 6000 system at 372M reads per sample. SNP recall calculated using FreeBayes v1.2.0.
Reduce bias in genomic interpretation
Coverage of challenging regions by the Invitrogen Collibri Physically Sheared (PS) DNA Library Prep Kits for high-throughput Illumina Systems is consistently even among low abundance (1 ng) and high abundance (1,000 ng) samples. This helps to ensure higher accuracy from high-throughput sequencing compared to older library prep methods from Illumina™, NEB™, or Kapa Biosystems™ kits. Unique Dual Indexes (UDIs) are available for 24 or 96 reactions to filter index-hopped reads on patterned flow cells.
Call variants with confidence
Consistently even GC coverage by the Collibri PS DNA Library Prep Kit for Illumina Systems and Collibri PCR-Free PS DNA Library Prep Kit for Illumina Systems enhances detection of single nucleotide polymorphisms (SNPs) and insertions/deletions (INDELs) with consistency from 1 to 1,000 ng of DNA. Sensitivity of detection is increased compared to older library prep methods.
Technical Note: Impact of GC bias on library preparation
Additional strategies are now available to minimize GC bias and achieve even genomic coverage.
Enhanced coverage of challenging regions, even from FFPE samples
Consistently even GC coverage produces higher coverage of difficult genomic regions, resulting in more even whole-genome coverage. Enhanced coverage of challenging genomic regions is consistent from 1 to 1,000 ng.
Figure 2. 100 ng of Horizon™ moderate damage FFPE DNA was converted into sequencing libraries using the manufacturer recommended protocols and sequenced on an Illumina NovaSeq 6000 system with a 2x150 bp read length. Resulting pass filter (PF) reads were normalized to 186 m reads.
Technical Note: Scaling library preparation and maintaining quality
Obtain consistent genomic coverage and variant detection in large sequencing projects which accommodate sample inputs from 1–1000 ng of DNA.
PCR is a Choice
Adaptor addition in the Collibri PS DNA Library Prep kits does not require PCR, and PCR-free protocols are available for 500–1,000 ng. Library yield, as measured by the Collibri Library Quantification Kit, from 500 ng of input DNA produces sufficient yield to complete quantification and multiple runs on an Illumina NovaSeq 6000 system.
Learn more about the PCR-free kit or the low input kit.
Figure 3. Coriell NA12878 DNA was converted into sequencing libraries using the manufacturer’s recommended protocols and concentration was measured using the Collibri Library Quantification kit.
Support across a faster workflow
Improve the efficiency of your lab with a faster library prep protocol and workflow support including DNA extraction, quantification, library preparation, and library quantification steps. Spend more time on science and less time on troubleshooting.
Click image to enlarge
Figure 4. Collibri PCR-Free PS DNA Library Prep protocols are completed in 1.5 hours, reducing total workflow time to ~6.5 hours.
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.
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DNA sequencing
Technical notes
All sequencing methods
Technical notes
RNA sequencing
Technical notes
For Research Use Only. Not for use in diagnostic procedures.