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Sample Handling01:02

Sample Handling

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Transportation of samples from the collection point to the laboratory, as well as storage and preservation techniques, are crucial for maintaining sample integrity and ensuring accurate and reliable test results.
Samples should be transported carefully from collection points to the laboratory. They should be properly sealed and clearly labeled to prevent cross-contamination. To preserve the sample integrity, optimal temperature conditions during transport are essential. This could involve using...
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Molecular Comparison of Gases, Liquids, and Solids02:26

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Particles in a solid are tightly packed together (fixed shape) and often arranged in a regular pattern; in a liquid, they are close together with no regular arrangement (no fixed shape); in a gas, they are far apart with no regular arrangement (no fixed shape). Particles in a solid vibrate about fixed positions (cannot flow) and do not generally move in relation to one another; in a liquid, they move past each other (can flow) but remain in essentially constant contact; in a gas, they move...
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Distribution reliability in electrical power systems is critical for ensuring an uninterrupted power supply to consumers at minimal cost. According to IEEE Standard Terms, reliability is the probability that a device will function without failure over a specified time period or amount of usage. For electric power distribution, this translates to maintaining continuous power supply and addressing customer concerns over power outages. Several indices, as defined by IEEE Standard 1366-2012, are...
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As with waves on a string, the speed of sound or a mechanical wave in a fluid depends on the fluid's elastic modulus and inertia. The two relevant physical quantities are the bulk modulus and the density of the material. Indeed, it turns out that the relationship between speed and the bulk modulus and density in fluids is the same as that between the speed and the Young's modulus and density in solids.
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High-Performance Liquid Chromatography: Introduction01:11

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High-performance liquid chromatography(HPLC), formerly referred to as High-pressure liquid chromatography, is a powerful technique used to separate, identify, and quantify components in complex mixtures. The term "high pressure" refers to using high pressure to push the liquid mobile phase through the tightly packed columns.
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Automated Robotic Liquid Handling Assembly of Modular DNA Devices
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High-throughput Quant-iT PicoGreen assay using an automated liquid handling system.

Kay Anantanawat1,2, Nicola Pitsch1, Caroline Fromont1,3

  • 1The Next Generation Sequencing Facility, Western Sydney University, Sydney, Australia.

Biotechniques
|May 17, 2019
PubMed
Summary

This study presents a high-throughput DNA quantification protocol using Quant-iT PicoGreen assay for next-generation sequencing (NGS) facilities. The optimized method significantly reduces reagent use and processing time for accurate nucleic acid concentration measurements.

Keywords:
DNA libraryPicoGreen assayhigh-throughputliquid handling systemsnext-generation sequencing

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genomics

Background:

  • Next-generation sequencing (NGS) workflows demand efficient sample processing.
  • Accurate nucleic acid quantification is crucial for downstream NGS library preparation.
  • Current methods can be time-consuming and reagent-intensive.

Purpose of the Study:

  • To develop a high-throughput protocol for DNA quantification.
  • To optimize the Quant-iT PicoGreen assay for large sample numbers.
  • To reduce reagent consumption and assay time in NGS facilities.

Main Methods:

  • A 384-well Quant-iT PicoGreen assay protocol was established.
  • An Eppendorf epMotion 5075 liquid handling system was utilized.
  • The protocol was designed for simultaneous duplicate measurement of DNA samples.

Main Results:

  • The protocol enables simultaneous measurement of 184 DNA samples in duplicate.
  • Assay completion time is reduced to 1 hour for 184 samples.
  • Reagent usage is decreased by 10-fold compared to standard protocols.
  • Overall processing time for 384 samples is reduced from 3 days to 3 hours.

Conclusions:

  • This high-throughput Quant-iT PicoGreen assay protocol significantly enhances efficiency in NGS facilities.
  • The optimized protocol offers substantial savings in both time and reagents.
  • It provides a robust solution for accurate nucleic acid concentration estimation in demanding research environments.