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NGSelect Amplicon

NGSelect Amplicon

Next-generation sequencing of amplicons of nearly any size

NGSelect Amplicon is an ideal service for scientists looking to perform NGS on PCR products unrestricted by amplicon size. With Illumina amplicon sequencing, extremely high coverage suitable for discovery of somatic mutations in complex DNA samples can be achieved. PacBio amplicon sequencing offers the chance to cover the whole length of the PCR products, eliminating the need for any assembly. Scientists can freely decide what part of the sequencing workflow they would like to outsource including PCR amplification and adapter ligation, sequencing or BioIT analysis. Proprietary protocols and years of NGS expertise help delivery high quality data with extraordinary fast delivery times.

Applications

The applications of NGSelect Amplicon are manifold. Some of the most typical uses of the service include:   

  • Screening of genetic variants in complex samples
  • Whole gene amplicon sequencing
  • Analysis of hard-to-sequence genomic regions, like highly repetitive areas, GC-rich areas, hairpin structures
  • Whole gene amplicon sequencing
  • Haplotype phasing for analysis of genetic co-variance
  • HLA allele typing
  • CRISPR sequencing and genome editing control
  • Sequencing of the 16S rRNA gene for microbial characterization

Highlights of NGSelect Amplicon

  • Sequencing of amplicons ranging from 100 bp to 6 kb in size
  • Large selection of Illumina sequencing options or PacBio sequencing options
  • Highly sensitive NGS workflows with reduced bias
  • Flexible data packages, starting from 5M reads
  • Customized combinations of sequencing modules: targeted amplicon sequencing, library preparation including PCR amplification and adapter ligation, as well as bioinformatics analysis
  • Multiplexing of up to 92 Illumina samples or 10 PacBio samples possible

 

NGSelect Amplicon is a uniquely designed service for next-generation sequencing (NGS) of amplicons or PCR products. The service offers researchers the choice of using only our sequencing capacities on the Illumina or PacBio platforms. Alternatively, customers can opt to combine the sequencing with other established segments of the sequencing process, including BioIT analysis. Amplicons as large as 6 kb can be reliably analyzed with this service. 

Starting Material

Library Type

 

Starting Material 

 

Illumina adapter ligation library

100 - 500 ng PCR product / amplicons (up to 100 µl, concentration > 1 ng/µl

Amplicon size range should ideally range from 100 bp to 570 bp (absolute min. 100 bp, absolute max. 650 bp) for HiSeq / MiSeq 

PacBio adapter ligation library

 

Amplicon size range of:

570 bp - 3 kb = 2 µg (up to 40 µl, concentration > 50 ng/µl)

3 kb - 6 kb = 4 µg (up to 40µl, concentration > 100 ng/µl)

 Table 1. Available genomic libraries and accepted starting material* for NGSelect - Amplicon

 

Specifications

Sequencing platforms

  • Illumina
  • PacBio

Data packages

Illumina data packages

  • starting from guaranteed 5M read pairs (10 million reads) per package

PacBio data packages

  • “Reads of Insert” ~15,000 reads (for long amplicon sizes of 570 bp to 3 kb)
  • “Reads of Insert” ~ 3,000 reads (for long amplicon sizes of 3 kb to 6 kb)

Bioinformatics analysis

Illumina

  • Merging of overlapping paired-end reads
  • Data sorting according to customer supplied tag

PacBio

  • “Reads of insert” analysis
  • Generation of consensus sequences from read inserts
  • Demultiplexing and sorting of reads (if applicable)

Delivery time

  • Starting from less than two weeks, varies depending on individual project aims

 

*For less starting material, please contact us.

Please note, that only S1-classified material is accepted for DNA isolation ordered online. More information about current classification of biological material can be found here (TRBA 466). Please contact us for further information on DNA isolation from material classified as S2.