Bioline NGS Library Preparation

Bioline NGS Library Preparation

The Bioline NGS Library Preparation Kits are designed to generate high-quality Next-Generation sequencing libraries suitable for sequencing on Illumina instruments. The success of Next Generation Sequencing is partly dependent upon the accurate processing of the input DNA. Library preparation from sheared DNA requires efficient processing during a series of molecular biology reactions and good recovery during the intermediate purification steps.

Faster Library Construction

By combining end repair, A-tailing and ligation in a single tube, the JetSeq NGS Library Preparation Kits offer a faster protocol with minimal effort resulting in a complete library construction in less than 3 hours. The elimination of purification steps minimizes sample loss, while optimization of the JetSeq buffer system ensures a high library yield.

Flexibility

A PCR and PCR-free kit for use with Illumina adapters and indexes, plus a complete kit that includes all of the components for multiplexing of up to 16 samples.

The PCR-free JetSeq ER & Ligation Kit contains all of the enzymes and buffers necessary for single tube end repair, A-tailing and ligation in one master mix and is recommended for Next Gen applications that do not require library PCR prior to sequencing.

The JetSeq Flex DNA Library Preparation Kit additionally contains the polymerase and buffer required for library amplification and is recommended for preparation of libraries from lower input material.

These kits do not contain adapters, but have been validated using standard, indexed Illumina adapters. They are also compatible with other adapters of similar design from other reputable oligonucleotide suppliers.

The JetSeq DNA Library Preparation Kit is a complete kit, containing the enzymes and buffers for end repair, A-tailing and ligation, the polymerase and buffer necessary for PCR amplification and the adapters for multiplexing of up to 16 libraries.

JetSeq™ Clean enables reaction clean-up, as well as a single- or double-sided size selection of libraries following the fragmentation, ligation and PCR steps in the next generation sequencing (NGS) library preparation workflow.

Higher Library adapter conversion efficiency

Ligation efficiency strongly influences library diversity and quality. The JetSeq kits are highly efficient at converting DNA into an adapter-ligated library, resulting in higher sequence coverage. The efficiency of the end repair and ligation steps can be quantified by a qPCR reaction of the adapter-ligated fragments prior to library amplification. By using adapter specific primers, the rate of conversion of input DNA to adapter-ligated fragments can be determined. JetSeq shows higher rates of conversion as compared to other commercially available kits.

Reduced Amplification bias

The GC content plays an important role a potential amplification bias. Different percentages of GC content in fragments can result relatively enriched or depleted fragments, resulting in misleading sequence data. Using a highly efficient polymerase together with an optimized buffer, the JetSeq amplification kits ensure uniform amplification of all fragments regardless of GC content. This ensures even coverage in subsequent sequencing reactions resulting in less sequencing per sample to reach a minimum level of coverage.

High-Quality Library Construction

Sequencing coverage describes the average number of reads that align to a known reference sequence and determines the confidence of variant discovery like SNPs, Indels and CNVs at particular positions. The enzymes in the JetSeq kits deliver exceptional sequence quality scores, giving greater coverage and reducing the possibility of sequencing gaps.

  • Low input: End-repair, A-tailing and ligation combined in the same tube, thereby eliminating cleanup steps and improving sample yield
  • Improved confidence: simpler protocol with fewer steps for reduced risk of sample loss and offering greater peace-of-mind
  • Increased speed: fast library preparation for reduced time to results and increased sample throughput
  • High-yield: PCR polymerase and buffer, developed specifically for library preparation, giving maximum yield of sequence-ready library DNA
  • Improved quality: optimized, high quality reagents result in reliable library preparation from even very challenging samples, providing maximum coverage
  • Flexibility: a PCR and PCR-free kit for use with Illumina adapters and indexes, plus a complete kit that includes all of the components for multiplexing of up to 16 samples.

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