Related Experiment Video
Updated: Jul 7, 2026
12:14
A Converging Strategy for the Generation of a Virtually Sequenced cDNA Library from Unreferenced Pacific Oysters
Published on: June 13, 2019
Production of a subtracted cDNA library.
1Brigham and Women's Hospital, Boston, Massachusetts, USA.
Current Protocols in Molecular Biology
|February 12, 2008
Summary
A subtracted cDNA library enriches for specific mRNA populations, useful for gene discovery when probes are unavailable. This method aids in identifying unique gene expression profiles between cell types.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Complete cDNA libraries can be inefficient for certain research goals.
- Identifying differentially expressed genes often requires specialized library construction.
- Subtracted cDNA libraries offer a targeted approach to gene discovery.
Purpose of the Study:
- To describe the utility and construction of subtracted cDNA libraries.
- To provide a protocol for generating subtracted cDNA libraries for gene isolation.
- To highlight applications in identifying specific mRNA populations.
Main Methods:
- Construction of a subtracted cDNA library by enriching for specific mRNA sequences.
- Utilizing a high-efficiency cloning vector, such as lambda gt10, for enhanced recombinant recovery.
- Application of subtraction protocols across various vector systems.
Main Results:
- Subtracted cDNA libraries contain clones corresponding to mRNAs present in one cell type but not another.
- This method facilitates the isolation of cDNA clones for specific mRNAs, especially when probes are unavailable.
- The protocol is optimized for high cloning efficiency and elimination of non-recombinants.
Conclusions:
- Subtracted cDNA libraries are valuable tools for isolating specific mRNA populations and aiding gene discovery.
- The protocol is adaptable and effective for identifying unique gene expression patterns.
- This technique streamlines the process of identifying novel genes or transcripts.
Related Concept Videos
02:35RACE - 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...
Since the...
03:21RNA-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...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
01:44Complementary DNA
Overview