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Next-generation Sequencing

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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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Genomics02:02

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Genomics is the science of genomes: it is the study of all the genetic material of an organism. In humans, the genome consists of information carried in 23 pairs of chromosomes in the nucleus, as well as mitochondrial DNA. In genomics, both coding and non-coding DNA is sequenced and analyzed. Genomics allows a better understanding of all living things, their evolution, and their diversity. It has a myriad of uses: for example, to build phylogenetic trees, to improve productivity and...
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Genomic Imprinting and Inheritance02:30

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Diploid organisms inherit genetic material through chromosomes from both parents. Copies of the same gene are known as alleles. In most cases, both alleles are simultaneously expressed and allow various cellular processes to function optimally. If one of the alleles is missing or mutated, the expression of the other allele can compensate; however, this is not true for all genes.
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Genome Size and the Evolution of New Genes03:21

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While every living organism has a genome of some kind (be it RNA, or DNA), there is considerable variation in the sizes of these blueprints. One major factor that impacts genome size is whether the organism is prokaryotic or eukaryotic. In prokaryotes, the genome contains little to no non-coding sequence, such that genes are tightly clustered in groups or operons sequentially along the chromosome. Conversely, the genes in eukaryotes are punctuated by long stretches of non-coding sequence.
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Cis-regulatory Sequences02:02

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Cis-regulatory sequences are short fragments of non-coding DNA that are present on the same chromosomes as the genes that they regulate. These fragments serve as binding sites for transcriptional regulators, proteins that are responsible for controlling gene transcription and differential gene expression across cell types in eukaryotes. Cis-regulatory sequences can be close to the gene of interest or thousands of bases away in the DNA sequence; however, those sequences that are further away are...
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Comparing Mitochondrial, Chloroplast, and Prokaryotic Genomes02:16

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The present-day mitochondrial and chloroplast genomes have retained some of the characteristics of their ancestral prokaryotes and also have acquired new attributes during their evolution within eukaryotic cells. Like prokaryotic genomes, mitochondrial and chloroplast genomes neither bind with histone-like proteins nor show complex packaging into chromosome-like structures, as observed in eukaryotes. Unlike mitotic cell divisions observed in eukaryotic cells, mitochondria and chloroplasts...
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Updated: Feb 14, 2026

Interactome-Seq: A Protocol for Domainome Library Construction, Validation and Selection by Phage Display and Next Generation Sequencing
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Whole Genome Library Construction for Next Generation Sequencing.

Jonathan J Keats1, Lori Cuyugan1, Jonathan Adkins1

  • 1Translational Genomics Research Institute (TGen), 445 N. Fifth Street, Phoenix, AZ, 85004, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 10, 2018
PubMed
Summary
This summary is machine-generated.

Whole genome sequencing (WGS) is a powerful genomics tool for biomedical research. This guide details WGS library preparation and quality control for Illumina sequencing, including methods for detecting structural variations.

Keywords:
Long insert whole genome sequencingNext generation sequencingShort insert whole genome sequencingWhole genome sequencing

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Area of Science:

  • Genomics
  • Molecular Biology
  • Bioinformatics

Background:

  • Whole genome sequencing (WGS) has become more accessible in biomedical research.
  • WGS is utilized for analyzing genomic DNA from individuals, families, and tumor samples.
  • Various WGS modalities exist, with a focus on library construction.

Purpose of the Study:

  • To describe wet lab library construction procedures for complex short insert WGS libraries.
  • To provide quality control measures for Illumina HiSeq2000 sequencing.
  • To present modifications for long insert WGS library construction for detecting structural alterations and copy number changes.

Main Methods:

  • Utilized the KAPA Hyper Prep Kit (Kapa Biosystems) for short insert WGS library construction.
  • Implemented quality control measures for the Illumina HiSeq2000 platform.
  • Adapted protocols for long insert WGS library construction.

Main Results:

  • Successfully constructed complex short insert WGS libraries.
  • Established quality control protocols for high-throughput sequencing.
  • Demonstrated feasibility of long insert WGS for structural variation analysis.

Conclusions:

  • The described methods provide a robust workflow for WGS library preparation and sequencing.
  • Quality control is essential for reliable WGS data generation.
  • Long insert WGS is valuable for detecting complex genomic rearrangements.