How does Illumina TruSeq work?
The TruSeq DNA Library Preparation Kits are used to prepare DNA libraries with insert sizes from 300–500 bp for single, paired-end, and indexed sequencing. An ‘A’- base is then added to the blunt ends of each strand, preparing them for ligation to the sequencing adapters (Figures 2C).
Is Illumina sequencing accurate?
Illumina Sequencing Quality Scores Illumina sequencing chemistry delivers high accuracy, with a vast majority of bases scoring Q30 and above. This level of accuracy is ideal for a range of sequencing applications, including clinical research.
What is PCR free WGS?
PCR Free Human Whole Genome Sequencing. Preparation of sequence-ready libraries without the need for PCR amplification presents advantages in data quality for human whole genome sequencing. These advantages include a marked reduction in base specific biases that are attributed with DNA polymerases.
What is TruSeq read?
The Read 1 primer is essentially the sequence of the TruSeq Universal Adapter, excluding of course the portion which binds to the flow cell. General Warning If you plan to create your own adapter sequences, indexes, and primers it is essential to KNOW WHAT YOU ARE DOING.
How does PCR Free library prep work?
The library prep process generates blunt end fragments from mechanically sheared DNA which are then size selected and biochemically prepared for a subsequent ligation process with Illumina adaptor indexes without the need for any PCR amplification.”
Does Illumina sequencing use primers?
The primer attaches to the DNA being sequenced. The DNA sequence is analysed base-by-base during Illumina sequencing, making it a highly accurate method. The sequence generated can then be aligned to a reference sequence, this looks for matches or changes in the sequenced DNA.
What are P5 and P7 adapters?
On each end, these adapter constructs have flow cell binding sites, P5 and P7, which allow the library fragment to attach to the flow cell surface. The P5 and P7 regions of single-stranded library fragments anneal to their complimentary oligos on the flowcell surface.
How is TruSeq used in the sequencing workflow?
With TruSeq technology, each step in this process is optimized to deliver accurate data to ensure the highest standard of quality for any research project. Platform accuracy describes the overall accuracy of the sequencing workflow, accounting for each step of the process, from sample preparation through variant discovery.
What are the steps in the sequencing workflow?
Accuracy Across the Illumina Sequencing Workflow. A typical sequencing workflow comprises sample/library preparation, cluster amplification, DNA sequencing, image analysis/base calling, read alignment, and variant discovery. If any of these steps generate poor results, the quality of the final data set is compromised.
Why is TruSeq used in Illumina library construction?
Library preparation is an especially crucial step; using TruSeq or other high-quality Illumina library construction technologies can help ensure high sample quality and accuracy for any research project.
How is TruSeq used to prepare small RNA samples?
The Illumina® TruSeq ™ Small RNA Sample Preparation protocol is used to prepare a variety of RNA species. The protocol takes advantage of the natural structure common to most known microRNA molecules.