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Related Concept Videos

RNA-seq03:21

RNA-seq

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 microarray-based...
RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

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 primer.
Since the...

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Cost-Efficient Transcriptomic-Based Drug Screening
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Cost-Efficient Transcriptomic-Based Drug Screening

Published on: February 23, 2024

Cost effective method for construction of high quality cDNA libraries.

Andreia Figueiredo1, Aladje Baldé, Hélia Cardoso

  • 1Unit of Molecular Biology and Plant Biotechnology-ICAT (Institute for Applied Science and Technology) Ed. ICAT, Campo Grande, 1749-016 Lisbon, Portugal. andreia.figueiredo@icat.fc.ul.pt

Biomolecular Engineering
|July 10, 2007
PubMed
Summary

This study presents a novel PCR-based method for synthesizing high-quality complementary DNA (cDNA). The technique enhances 5' end representation and saves time and resources in molecular biology research.

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CRISPR-Based Modular Assembly for High-Throughput Construction of a UAS-cDNA/ORF Plasmid Library
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CRISPR-Based Modular Assembly for High-Throughput Construction of a UAS-cDNA/ORF Plasmid Library

Published on: May 17, 2024

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genomics

Background:

  • Complementary DNA (cDNA) synthesis is crucial for gene expression studies.
  • Traditional cDNA synthesis methods can be inefficient, especially with limited starting material.
  • Current protocols often involve multiple laborious steps like digestion and subcloning.

Purpose of the Study:

  • To develop an improved PCR-based method for high-quality cDNA synthesis.
  • To enhance 5' end representation in cDNA libraries.
  • To streamline the cDNA synthesis process, reducing time and cost.

Main Methods:

  • Utilized a PCR-based approach.
  • Integrated reverse transcriptase template switching.
  • Employed assisted recombination cloning techniques.

Main Results:

  • Achieved high-quality cDNA synthesis.
  • Enabled 5' end enrichment.
  • Successfully synthesized cDNA from limiting starting material.
  • Eliminated the need for digestion and subcloning steps.
  • Demonstrated substantial time and cost savings.

Conclusions:

  • The described method offers an efficient and cost-effective alternative for cDNA synthesis.
  • This technique is particularly advantageous for applications requiring 5' end enrichment or working with scarce RNA samples.
  • The streamlined protocol simplifies molecular biology workflows.