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Related Concept Videos

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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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DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
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RNA-seq03:21

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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
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RACE - Rapid Amplification of cDNA Ends02:35

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Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific...
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
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Updated: Sep 26, 2025

Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites
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Progress on methods for acquiring flanking genomic sequence.

Ji-Ming Xu1, Jian-Shu Zhu1, Meng-Zhen Li1

  • 1Institute of Plant Biology, College of Life Science, Zhejiang University, Hangzhou 310058, China.

Yi Chuan = Hereditas
|April 19, 2022
PubMed
Summary
This summary is machine-generated.

Flanking genomic sequences are crucial for understanding gene regulation and chromosome structure. This review compares various methods for acquiring these important DNA sequences, aiding genomics research.

Keywords:
flanking sequenceinverse PCRligation-mediated PCRplasmid rescuesemi-random primer PCRwhole-genome resequencing

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Area of Science:

  • Genomics
  • Molecular Biology
  • Biotechnology

Background:

  • Flanking genomic sequences provide critical information on candidate genes, transcriptional regulation, chromosome structure, and biosafety.
  • These sequences are vital for structural and functional genomics research, including applications in transgenic organism identification and safety management.

Purpose of the Study:

  • To review and compare diverse methods for obtaining flanking genomic sequences.
  • To discuss the principles and recent advancements in flanking sequence acquisition technologies.

Main Methods:

  • Plasmid rescue
  • Inverse PCR (Polymerase Chain Reaction)
  • Ligation-mediated PCR
  • Semi-random primer PCR
  • Whole-genome resequencing

Main Results:

  • The review summarizes established and emerging techniques for flanking sequence acquisition.
  • A comparative analysis of the principles and research progress of each method is presented.

Conclusions:

  • Various molecular biology techniques enable the acquisition of flanking genomic sequences.
  • Understanding these methods is essential for advancing genomics research and its applications.