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Related Concept Videos

DNA-only Transposons02:57

DNA-only Transposons

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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
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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. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
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Transposons01:24

Transposons

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Transposons, or "jumping genes," are small mobile genetic elements (MGEs) that range from 700 to 40,000 base pairs in length. They are found in all organisms and can move within the same chromosome or transfer to different chromosomes. In some cases, transposons can also jump between different host DNA molecules, such as plasmids or viruses, contributing to genetic variability.Barbara McClintock first discovered these mobile genetic elements in the 1940s while studying maize genetics, and she...
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Overview of Transposition and Recombination02:13

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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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Related Experiment Video

Updated: Oct 6, 2025

Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing
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Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing

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dCITI-Seq: droplet combinational indexed transposon insertion sequencing.

Jing Tu1, Yi Qiao2, Zheyun Xu2

  • 1State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, China. jtu@seu.edu.cn.

Analytical and Bioanalytical Chemistry
|January 19, 2022
PubMed
Summary

We developed droplet combinational indexed transposon insertion sequencing (dCITI-Seq) for scalable library construction. This method enhances indexing capacity and reduces errors for high-throughput sequencing applications.

Keywords:
Combinational indexingDroplet mergingHigh-throughput sequencingTn5 transposition

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • High-throughput sequencing demands efficient large-scale barcoding and library construction.
  • Existing methods face challenges in scalability and accuracy.

Purpose of the Study:

  • To introduce droplet combinational indexed transposon insertion sequencing (dCITI-Seq) as an improved method for library preparation.
  • To address limitations in indexing capacity and index crosstalk in large-scale sequencing.

Main Methods:

  • Utilized a droplet pairing and merging platform for random combination of two adaptor sets with barcodes.
  • Employed massively parallel transposition for direct insertion of index-containing adaptors.
  • Developed custom bioinformatics pipeline for data processing.

Main Results:

  • dCITI-Seq demonstrated enlarged indexing capacity and reduced index crosstalk.
  • Observed a lower GC base preference compared to conventional in-tube transposition methods.
  • Successfully applied dCITI-Seq to large-scale single-cell sequencing.

Conclusions:

  • dCITI-Seq offers a robust and scalable solution for sequencing library construction.
  • The method improves accuracy and efficiency for high-throughput genomics.
  • Potential for broad application in large-scale single-cell sequencing studies.