EasySeq NGS smMIP based capture

EasySeq NGS Targeted Capture Molecular Inversion Probes

Brand introduction

Technology of EasySeq NGS Targeted Capute

Using sequence capturing with single molecule Molecular Inversion Probes (smMIP), followed by a barcoding amplification step, the system enables sequencing of the targeted regions in a very simple, fast and robust workflow, and is enabling coverage with double probe tiling and independent reading of both strands.

The kits contain all reagents for a straightforward, one-step capturing and subsequently sample tagging for NGS sequencing. The resulting libraries are compatible with illumina® sequencing platforms.

EasySeq NGS RC-PCR

EasySeq NGS Reverse Complement PCR

RC-PCR

Contamination Concerns?

Conventional Amplicon based methods for NGS target enrichment involve two PCR steps – one to amplify the regions of interest followed by a second PCR reaction to add adapter sequences for NGS. While these methods are well established there is always a high risk of cross contamination or carry-over from first to second rounds of amplification

Legal Notice:
RC-PCR is IP protected.
US Patent / WO Patent / EP Patent

Exclusive license granted to NimaGen B.V., Nijmegen, The Netherlands
by SALISBURY NHS FOUNDATION TRUST (SALISBURY, WILTSHIRE, GB)

It is strictly prohibited to use, produce, distribute, import, or sell RC-PCR based methods or kits without the patent owner’s written consent.

How it Works

Easy Handling

Just one Pre-PCR reaction setup, quick, easy and highly suited for automation

RC-PCR

Product generation is more target specific

Traditional PCR reactions have high concentrations of primer at the beginning of the reaction when the template availability is low. 

In a RC-PCR the target specific primer is synthesized during the reaction and consequently more aligned with the target availability reducing the chance for dimerization and off target binding.

Multiplex Capabilities

Predesigned and custom panels available targeting up to 100 amplicons in a single closed tube reaction.

Solutions available and in development: 
  1. Human Sample tracking and ID kit for WGS/WES QC
  2. Forensic Identification Kits
  3. Crop Genotyping assays (GreenoType)
  4. GMO detection kit
  5. Meat detection kit (including speciate)
  6. Pharmacogenetics (PGX)
  7. Circulating Tumour DNA kit
  8. Detection of aneuploidies

Magnet Plate

Alpaqua Magnet Plate

Portfolio

Alpaqua magnet for automation and manual use

Benefits of Ring Magnets

One dedicated magnet for each well means fewer edge effects and greater uniformity of results, improving subsequent resuspension and elution and offering near universal microplate compatibility.

Patented Spring Cushion Technology

Magnet plates with integrated spring cushion technology give way when tips come in contact with a well bottom, thus compensating for physical tolerances between labware and pipettors that can compromise precision aspiration.

Let us find your perfect match

Not sure which magnet plate is best for your specific application? We can help you find the best solution.

Are you looking for speed? Low elution volumes? Or using exotic PCR plates? Feel free to contact us with the requirements for your workflow!

covid19

EasySeq™ RC-PCR SARS CoV-2 (novel coronavirus) Whole Genome Sequencing

Generate Whole Genome Sequencing data from cDNA, derived from Covid-19 positive tested patients.

Introduction

Info

The analysis and study of viral genomes is an essential epidemiological requirement to track the evolution of a virus, the origin of strains and their transmission during any epidemic or pandemic. Targeted Next Generation Sequencing (NGS) is a valuable tool for both detection and classification of viral genomes. The 2019 outbreak of the novel Coronavirus strain (SARS-CoV-2) is obviously unprecedented with over 26 million confirmed cases of the disease associated with SARS-CoV-2 (COVID-19) and sadly approaching 1 million of lost lives in a relatively short period of time. Therefore, the ability for public health professionals and researchers to monitor situations in the most efficient and streamlined manner possible is essential. As is the reassurance that testing methods are secure, and results are accurate.

RC-PCR, the next revolution in NGS Library Prep

– More efficient ­library prep: Up to 80% less hands-on time
– Combined Amplification and Indexing with RC-PCR
– Less Handling, Less Risk, Greater Sample Safety
– Easily adapted for Automation
– Single Click Analysis with virSEAK (JSI Medical Systems)

With the NimaGen EasySeq™ RC-PCR Library Preparation method the implementation of an NGS driven viral surveillance strategy could not be simpler or safer. Utilising patented RC-PCR technology, sequence ready libraries are generated from cDNA covering the entire 29.9Kb SARS-CoV-2 Genome with minimal hands-on time and just a single PCR for both amplification and indexing. Tiled RC-PCR primers and probes are provided in tubes, as well as 2 breakable 96 well plates which include UDI Indexes. Just add cDNA and the RC-PCR mastermix provided in the kit and load on to your PCR instrument. Post PCR all samples can be combined for pooled cleanup with the included Ampliclean Magnetic Bead Cleanup Kit and following standard quantification and QC checks your libraries are ready to sequence.

Video

Performance

Thanks to the improved reaction kinetics, intrinsic in the RC-PCR method, in combination with the well designed and balanced probe pools, the kit delivers clean data. Also, this allows for well balanced read distributions in order to make efficient use of the sequencer’s capacity. The method delivers a coverage of ~99% of the Covid Genome, and an on-target percentage of >90%. Thanks to the easy workflow, with just a few pipetting steps, this kit is the ideal solution for any virology lab, even without NGS experience. Our support team will get you up and running in no time.

Data Analysis

VirSEAK Analysis software

JSI medical systems GmbH in Germany developed a dedicated software tool for analyzing NGS data from the SARS-CoV-2 virus, with all needed settings ready installed.

Please contact JSI Medical Systems directly to learn about this software

Global GISAID data integrated

 

After the sequencing run ends, the generated fastq files can be imported into the JSI virSEAK software tool. virSEAK will automatically create a consensus sequence and alignment to the reference genome. It will show the claid where the sample belongs to and generate a .csv report of the mutations. Data will be compared to global databases (like GISAID) and you will be shown the subtype frequencies and distribution per country, as well as the evolution of the strain.

Results can be uploaded to the GISAID database with a single mouse-click in order to share the results with the global Covid community. It also provides an exportable consensus sequence for a more detailed analysis in local healthcare systems. For instance when investigating patient clusters or re-infections.

Product

EasySeq™ RC-PCR SARS CoV-2 (novel coronavirus) Whole Genome Sequencing kit (96 reactions). 

SKU: RC-COV096

 

 
Format

Generate Whole Genome Sequencing data from cDNA, derived from Covid-19 positive tested patients.

Compatibility

Compatible with any low- and mid throughput illumina® NGS sequencer of the latest generations: iSeq®, MiniSeq®, MiSeq®, NextSeq®, in combination with 2×150 bp paired-end read chemistry.

Product Information Variants Cat.
EasySeq™ RC-PCR SARS CoV-2 (novel coronavirus) Whole Genome Sequencing kit (96 reactions). 96 reactions SKU: RC-COV096
PCR 2x Hotstart HiFi Mastermix, compatible with RC-PCR assays, 2.2 mL 2 Tubes of 1.1 mL  each SKU: MMHS-25

Next Generation Sequencing

Next Generation Sequencing

Portfolio

Targeted library prep solutions

NimaGen offers different library prep kits for Illumina. All kits are based on our proprietary Reverse-Complement PCR or on the single-molecule Molecular Inversion Probe (smMIP) methods. 

Reverse Complement PCR (RC-PCR) based assays:

1. EasySeq NGS RC-PCR

Reverse-Complement PCR: The only one-step library prep method on the market.

  • ➼ The safest
  • ➼ The fastest
  • ➼ The simplest

2. EasySeq NGS smMIP based capture

Single-molecule targeted capturing, highly quantitative due to Unique Molecular Identifiers (UMI).

Sanger Sequencing

Sanger Sequencing

Introduction

Sanger method of DNA sequencing

Sanger sequencing is a method of DNA sequencing, based on the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication, developed by Frederick Sanger and colleagues in 1977. Since the introduction of massive parallel sequencing (NGS), the Sanger method remains in wide use, for smaller-scale projects, validation of NGS results and for obtaining especially long contiguous DNA sequence reads.

Fluorescent DNA Fragment Analysis

Fluorescent DNA fragment analysis is one of the most useful methods in molecular biology. The method measures the relative size of DNA fragments with a very high resolution and reproducibility, by capillary electrophoresis (CE) of fluorescent labelled DNA fragments on an automated DNA CE Genetic Analyzer, using internal fluorescent size standards.

Portfolio

NimaGen offers a complete portfolio of high-quality and cost-effective reagents for Applied Biosystems capillary electrophoresis based Genetic Analyzers, like the 310, 3100 series, 3130 series, 3500 series, 3730 series and SeqStudio.