What is PCR-free sequencing?

What is PCR-free sequencing?

By eliminating PCR amplification steps, the PCR-free protocol significantly reduces typical PCR-induced bias and provides detailed sequence information for traditionally challenging regions of the genome.

What is Illumina TruSeq?

TruSeq DNA Library Preparation Kits are a simple, cost-effective solution for generating high-quality libraries, compatible with unparalleled Illumina sequencing output. TruSeq library preparation has become the industry’s most trusted and widely adopted library preparation method for next-generation sequencing (NGS).

What is Illumina PCR?

Illumina DNA PCR-Free Prep A high-performing, fast, and integrated workflow for sensitive applications such as human whole-genome sequencing.

What is the difference between TruSeq and Nextera?

Illumina TruSeq and Nextera technologies are identical in many characteristics, except that Nextera uses transposomes for fragmentation, whereas TruSeq fragments the DNA by ultrasonication. The Agilent technology uses RNA molecules as probes, whereas all the other technologies use DNA as probe molecules.

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.”

What is the difference between PCR and whole genome sequencing?

Polymerase Chain Reaction (PCR) is the process which creates a large number of copies of a DNA fragment. DNA sequencing is the technique which results in the precise order of the nucleotides of a given DNA fragment. This is the key difference between PCR and DNA sequencing.

What are the steps to Illumina sequencing?

Figure 3: Next-Generation Sequencing Chemistry Overview—Illumina NGS includes four steps: (A) library preparation, (B) cluster generation,(C) sequencing, and (D) alignment and data analysis.

Does Illumina require PCR?

The Illumina Lysis Kit is required only if you are processing blood samples with Illumina DNA PCR-Free Prep. The Illumina Free Adapter Blocking Reagent is optional. It is used to reduce index hopping levels. Sign in to see pricing and favorite products.

How do you create a PCR protocol?

A standard polymerase chain reaction (PCR) setup consists of four steps:

1. Add required reagents or mastermix and template to PCR tubes.
2. Mix and centrifuge.
3. Amplify per thermo cycler and primer parameters.
4. Evaluate amplified DNA by agarose gel electrophoresis followed by ethidium bromide staining.

What is P5 and P7 in sequencing?

The P5 region is cleaved, resulting in clusters containing only fragments which are attached by the P7 region. This ensures that all copies are sequenced in the same direction. The sequencing primer anneals to the P5 end of the fragment, and begins the sequencing by synthesis process.

What is P5 and P7?

P5 and P7 are names given by Illumina to the oligo sequences that bind to the flow cell. Source publication. Multiplexed Illumina sequencing libraries from picogram quantities of DNA.

What is Illumina DNA PCR free prep?

Illumina DNA PCR-Free Prep A high-performing, fast, and integrated workflow for sensitive applications such as human whole-genome sequencing. TruSeq DNA Nano Generate whole-genome sequencing libraries and efficiently interrogate samples with limited available DNA.

How does TruSeq DNA PCR-free improve library preparation?

Bead-based size selection shortens the workflow. In tandem with Illumina sequencing systems, TruSeq DNA PCR-Free provides a range of enhancements to a widely adopted library preparation workflow. Access flexible throughput options TruSeq DNA PCR-Free with Single Indexes supports 24-sample manual processing for low-throughput (LT) studies.

What is the throughput of TruSeq DNA PCR?

Access flexible throughput options TruSeq DNA PCR-Free with Single Indexes supports 24-sample manual processing for low-throughput (LT) studies. TruSeq DNA PCR-Free with 96 CD Indexes supports 96-sample processing for high-throughput (HT) studies, and can be automated on liquid-handling robots (or processed manually).

Why choose Illumina Genome sequencing?

The workflows are tunable to a variety of read lengths and are supported on all Illumina sequencing instruments. This permits the researcher to tailor each run to the needs of the experiment. Gain deep insight into the genome PCR-free means reduced library bias and gaps.