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The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
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Sequences are fundamental mathematical objects consisting of ordered lists of numbers that follow a specific rule or pattern. Sequences are critical in various mathematical concepts, including calculus, series, and number theory. They can model real-world phenomena such as population growth, financial investments, and physical processes like the diminishing height of a bouncing ball.Each number in a sequence is referred to as a term. Typically, the terms are denoted as a1, a2, a3,…, where...
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An arithmetic sequence is a structured arrangement of numbers where each term is derived by adding a constant value, known as the common difference, to the previous term. This consistent pattern allows for the efficient computation of any term within the sequence as well as the cumulative sum of multiple terms. The formula for finding the nth term of an arithmetic sequence is:Here, aₙ represents the nth term of the sequence, a is the first term, d is the common difference, and n is the...
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Updated: Feb 15, 2026

RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing
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Rapid and Easy Protocol for Quantification of Next-Generation Sequencing Libraries.

Steve F C Hawkins1, Paul C Guest2

  • 1Bioline Reagents Limited, Unit 16, The Edge Business Centre, London, UK. shawkins@bioline.com.

Methods in Molecular Biology (Clifton, N.J.)
|January 31, 2018
PubMed
Summary

Accurate DNA quantification is essential for next-generation sequencing (NGS) library preparation. A new real-time quantitative PCR (qPCR) protocol offers precise DNA quantification, improving NGS data quality and enabling studies on nutritional programming and medical disorders.

Keywords:
Next-generation sequencingPrimersRNATemplateqPCR

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Next-generation sequencing (NGS) has revolutionized DNA sequencing efficiency.
  • Accurate quantification of NGS libraries is critical for reliable sequencing data, especially on Illumina platforms.
  • Current spectrophotometric and fluorometric methods lack the precision of quantitative PCR (qPCR).

Purpose of the Study:

  • To present a real-time quantitative PCR (qPCR) protocol for precise DNA quantification.
  • To provide a method applicable to next-generation sequencing (NGS) library preparation.
  • To highlight the utility of accurate DNA quantification in various research fields.

Main Methods:

  • Development and application of a real-time quantitative PCR (qPCR) protocol.
  • Utilizing qPCR for precise DNA quantification in the context of NGS library preparation.
  • Comparison of qPCR with traditional spectrophotometric and fluorometric quantification methods.

Main Results:

  • The presented qPCR protocol enables exact DNA quantification.
  • This method ensures high-quality data output from Illumina sequencing platforms.
  • The protocol is suitable for diverse applications, including studies on nutritional programming.

Conclusions:

  • Real-time qPCR offers superior accuracy for DNA quantification compared to physical property analysis methods.
  • Implementing this qPCR protocol can significantly enhance the reliability of NGS experiments.
  • Accurate NGS library quantification is vital for advancing research in areas like medical disorders linked to nutritional programming.