NGSelect Amplicons

NGSelect Amplicon

Short-Read Amplicon Sequencing using Illumina.

Short-read amplicon sequencing is a uniquely designed service for scientists looking to perform NGS on clonal or complex PCR products using either Illumina´s MiSeq or NovaSeq. A complex amplicon is a PCR product characterized by high genetic variability, structural variations, or mixed populations, such as those found in somatic alleles or 16S rRNA studies, making sequencing and analysis more challenging.

Take advantage of great prices by reducing sequencing costs with inline tags for a second-level multiplexing or opt for smaller read packages for more flexibility.

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Highlights

Designed to resolve complex amplicon mixtures
Quality control assessment
Flexible data packages, starting from 60 k read pairs up to full flow cells
Multiplexing flexibility enables a more cost-effective assay
Services are performed in ISO17025 certified labs
Wide range of bioinformatics services & interactive analysis reports
Data transfer via secure FTP

Overview

Size	150 – 270 bp	280 – 570 bp
Data package	5M read pairs	60k read pairs*
Sequencing technology	NovaSeq, 150 PE	MiSeq, 2x300bp
Quality control	Yes, quantity and size control	Yes, quantity control
TruSeq adapter sequence**	No	Yes Primer design and ordering tool available at: NGSgrade Oligos for MiSeq (eurofinsgenomics.eu)
Inline tags**	Up to 192 inline tags per sample: designed by our ordering tool NGS Adapter Ligation Oligos (eurofinsgenomics.eu)	No
Input material	25 µL purified PCR products Qubit quantified (> 4 ng/µl)	25 µL purified PCR products Qubit quantified (> 1 ng/µl)

*Full MiSeq flow cell sequencing options are available using 2x250 bp and 2 x 300 bp sequencing mode as well, please contact our customer support team.

** please refer to specifications below

Application & Services

Amplicon size	150-270 bp	280-570 bp
Application	CRISPR Check Analysis 16S Microbiome Analysis Unique Sequence Analysis Consensus Sequence Analysis Variant Analysis – Germline Variant Analysis - Somatic	CRISPR Check Analysis 16S Microbiome Analysis Unique Sequence Analysis Consensus Sequence Analysis Variant Analysis - Germline
Application	Trimming & Merging Inline Tag Sorting Sequence Cleaning Host Removal	Trimming & Merging Sequence Cleaning Host Removal

Specifications

A. NGSelect Amplicons on NovaSeq (150 - 270 bp)

Sequence your 150-270 bp long amplicons and benefit from the ability to pool up to 192 amplicons in a single sample using inline tags. Simply provide your amplicons with or without inline tags, but please avoid any TruSeq or Nextera adapter sequences.

You are unsure on how to add the inline tag sequences? Please check our tool: NGS Adapter Ligation Oligos (eurofinsgenomics.eu). Make sure to provide inline tag information in the sample submission form.

Achieve extremely high coverage, ideal for discovering somatic mutations in complex DNA samples. Our proprietary protocols and extensive NGS expertise ensure high-quality data with fast delivery times.

You can choose between multiples of guaranteed 5 M read pair packages (+/- 5%) for NovaSeq (2 x 150 bp)

Starting material

Amplicon size: 150 - 270 bp
Sample Type: Purified PCR products
Purity (OD260/280): 1.8-2.0
Resuspension Buffer: Water, EB, or low TE (<0.1 mM EDTA)
Format: barcode labelled 1.5 ml safe-lock tubes.
Shipment Method: Room temperature/ice/dry ice

Volume, Amount and Concentration depend on the product (Standard or Premium). All details can be found on our sample submission page >>

B. Amplicons on MiSeq (280 - 570 bp)

Sequence your 280-570 bp long amplicons and take advantage of our small read packages. Simply provide your amplicons with TruSeq adapter sequences, and we will handle the rest.

The adapter sequences are:

Forward: 5’-ACACTCTTTCCCTACACGACGCTCTTCCGATCT-3’;

Reverse: 5’-GACTGGAGTTCAGACGTGTGCTCTTCCGATCT-3’

You are unsure on how to add these sequences? Please check our tool: NGSgrade Oligos for MiSeq (eurofinsgenomics.eu).

Our services include indexing all samples and sequencing on the MiSeq platform. You can choose between guaranteed 60k read pairs (+/- 3%) data output, as well as full MiSeq runs (2 x 300 bp or 2 x 250 bp)

Starting material

Amplicon size (not including the universal adapters): 280 - 570 bp
Sample Type: Purified PCR products
Purity (OD260/280): 1.8-2.0
Volume: 25µL
Concentration: >1 ng/µl
Minimum Amount: 25 ng
Resuspension Buffer: Water, EB, or low TE (<0.1 mM EDTA)
Format: barcode labelled 1.5 ml safe-lock tubes; or for > 48 samples in Eppendorf twin.tec PCR Plate 96, full-skirted, leave position G12 & H12 empty
Shipment Method: Room temperature/ice/dry ice