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Quantification and normalization

FROM ABSORBANCE- AND FLUORESCENCE READING TO INTEGRATED FRAGEMENT ANALYSIS

Whether you want to quantify just a few DNA or RNA samples, or need fully integrated quality control for your automated workflows, Tecan has the right solution to meet your needs.

Quantification and normalization of nucleic acids following nucleic acid extraction or as part of NGS workflows avoids poor quality results or downstream failures due to insufficient or contaminated samples, saving time and lowering costs.

Related products:  Frida Reader™  |Infinite® 200 PRO  |   TouchTools™  |   Freedom EVO®  |   Fluent®  |   Consumables




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Tab 01 / Stand-alone DNA and RNA quantification
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Sensitive and fast quantification

The Infinite 200 PRO multimode reader family offers absorbance and high sensitivity fluorescence reading modes in one compact instrument.

It combines the high sensitivity of fluorescence assays with the convenience of label-free UV analysis for formats up to 384-well plates.

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High performance DNA quantification
and QC

The Infinite M Nano absorbance microplate reader, one of six new tailored configurations based on the acclaimed Infinite 200 PRO, offers rapid quantification of DNA and RNA with full format flexibility, from cuvettes to 384-well plates.

Patented Quad4 Monochromators™ technology gives you total confidence in sample quality, based on complete spectral analysis from 230-1000 nm.

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Accurate small volume analysis

The patented NanoQuant Plate™ represents the most accurate and reproducible low volume nucleic acid quantification tool on the market, for samples as small as 2 µl.

Its high precision quartz optic guarantees excellent performance for the simultaneous measurement of up to 16 samples, without the need for calibration. Compatible with any Tecan multimode reader, this powerful tool is perfect for DNA/RNA quantification and quality control in small volume samples.

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Tab 02 / Integrated quant. and normalization
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Frida Reader

The Frida Reader module for Fluent workstations allows quantifying nucleic acids without sample loss. Designed to work in combination with the Air Flexible Channel Arm™ (AirFCA) of the Fluent Automation Workstation, this patented approach performs UV absorbance-based quantification and purity assessment in a hanging drop, avoiding the consumption of rare and precious samples following nucleic acid purification (NAP). It is ideal for precious or low volume samples, as the sample is aspirated back into the tip and can directly be used for further processing, such as normalization.

Download more information on the Frida Reader

Infinite 200 PRO

DNA quantification can be achieved by measuring an aliquot with a fluorescent dye, such as PicoGreen® or QuantiFluor™, or by measuring the absorbance of the entire DNA sample.

DNA quantification and normalization workflows typically include the following steps:

  • Set-up of quantification assay
  • Determination of the DNA concentration in a reader
  • Calculation of a dilution scheme to achieve the target concentration for each well
  • Dilution of the DNA samples to the desired concentration

The Infinite 200 PRO can be integrated on the workdeck, on a side extension or below the workdeck, ensuring an optimal fit into your genomics workflow.

Straightforward integration of an Infinite reader on or below the workdeck of a Fluent platform.

Normalization workflow

The Freedom EVOware® Normalization Wizard is a software component that efficiently automates pipetting steps for the quantification and normalization of nucleic acid samples.

Samples of varying concentrations are diluted to a uniform concentration across the entire microplate. This enables Freedom EVO workstations to perform all the pipetting and microplate handling steps required for sample normalization tasks.

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FluentControl™ smart commands

Save time with a few simple commands for quantification and normalization that execute complex tasks.

  • Quantify DNA by calculating concentrations from standard curves
  • Normalization command sets up concentration-based pipetting scheme
  • New Dilution command to execute all necessary pipetting steps
  • FluentControl comes with example scripts to aid development
Read more




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Tab 03 / Integrated fragment analysis
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Fragment Analyzer INFINITY™
integrated nucleic acid quality control

Fragment analysis

Traditionally, quality analysis of DNA fragments is performed by agarose gel electrophoresis, which is both time consuming and labor intensive.

The Fragment Analyzer INFINITY (FAI) from Advanced Analytical offers automated multichannel capillary electrophoresis, providing high resolution separation of 96 samples in parallel.

It can accurately size and quantify nucleic acid fragments and smears from 50bp to 50kb, and can hold two different gel reagents for seamless switching between applications and sample types.

Over 20 reagent kits are available – each optimized for the analysis of different fragment lengths – for:

Integration of the FAI onto a Freedom EVO® liquid handling platform enables incorporation of important quality control checkpoints into automated genomics workflows.

This allows longer walkaway times and ensures that DNA fragments meet the quality control criteria necessary for subsequent sequencing or other downstream analyses.

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Integration

Fully automated DNA quality control with Fragment Analyzer INFINITY integration on the Freedom EVO

 

Tecan Labwerx™ can integrate the FAI onto the right side of a Freedom EVO workstation, allowing it to be accessed by the system's Robotic Manipulator Arm. This configuration provides easy access to the FAI side panel to refill gels and conditioning liquids, as well as to empty the waste bottle.

Please see this technical note for further information on hardware and software integration.




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Tab 04 / Literature
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Brochures

Accurate nucleic acid quantification and quality control without sample loss

Module for Fluent® automation workstation specifications and typical performance values

Software to help you normalize your DNA samples to uniform concentration

Application Notes

Nucleic acid quantification and normalization is required in most genomics workflows, most commonly after the initial nucleic acid purification step, but consumes precious sample and can be tedious when performed manually. The versatility of the classical UV absorbance-based technique still makes it the ‘go to’ method for many applications, as it allows the concentration of all DNA and RNA types to be measured (based on the specific empirical extinction coefficient for each nucleic acid type) and provides simultaneous assessment of samples for impurities that could impair downstream applications.

The Frida Reader is an add-on module for the Fluent Automation Workstation that revolutionizes the way quantification and normalization are performed. It allows samples to be measured in a hanging drop at the end of a disposable pipette tip, completely eliminating sample loss for nucleic acid quantification.

DNA quantification and normalization are critical steps for a wide range of genomics applications, such as next generation sequencing and quantitative PCR. Due to the limitations of classical absorbance techniques, a number of more sensitive methods for quantification of double stranded DNA (dsDNA) have been developed over the years. Fluorescence-based techniques have proved popular with researchers, as the introduction of fluorescent dsDNA-binding dyes with minimal binding to single stranded DNA (ssDNA) and RNA allows selective quantification of even small amounts of dsDNA.

Applicability of the NanoQuant PlateTM for Fluorescence Measurements.

Absorbance measurement on the Infinite® M200 microplate reader using the NanoQuant Plate™.

NanoQuant Plate™ for use on Infinite® F200 and Infinite® M200

Quantification and normalization of double stranded DNA with minimal set-up time.

Fast and simple DNA quantification and normalization

Technical Notes

Comparing absorbance measurements between the Quad4 Monochromators™-based Infinite® M200 PRO and a multimode reader using photodiode array technology.

NanoQuant Plate™ for use of Infinite® 200 NanoQuant.

Automated QC analysis for NGS Library Preparation and CRISPR/CAS9 Gene Editing

User Guides

The Frida Reader for Fluent offers UV-based concentration and purity measurements of nucleic acid samples in a hanging drop at the end of a disposable tip. This patented method is completely free of sample loss, as the hanging drop is aspirated back into the tip, and can be used for further processing (e.g. a normalization). No additional preparation steps, labware or reagents are required for the measurement, which offers precision and accuracy comparable to a reference reader over a range of 2 to 1,000 ng/μl.

For research use only. Not for use in clinical diagnostic procedures.