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The use of multiple displacement amplification to amplify complex DNA libraries
Melissa J Fullwood1, Jack J S Tan, Patrick W P Ng
1Genome Institute of Singapore, Agency for Science, Technology and Research (A*STAR), 60 Biopolis Street, Genome #02-01, Singapore.
Nucleic Acids Research
|February 21, 2008
Summary
This study introduces a novel method for DNA amplification, combining bacterial selection with multiple displacement amplification (MDA). This approach efficiently selects single DNA clones, reducing bias and improving scalability for complex genomic and cDNA library sequencing.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Bacterial propagation is standard for DNA sequencing library amplification but is time-consuming and prone to bias.
- Multiple displacement amplification (MDA) amplifies DNA in vitro but lacks a mechanism to remove unwanted ligation multimers.
Purpose of the Study:
- To develop an unbiased and efficient method for amplifying complex DNA libraries.
- To overcome the limitations of traditional bacterial propagation and MDA for DNA library preparation.
Main Methods:
- A novel technique involving brief bacterial introduction of ligation reactions to select single DNA insert clones.
- Subsequent amplification of selected clones using multiple displacement amplification (MDA).
- Application of the method to a Gene Identification Signatures with Paired-End diTags (GIS-PET) library.
Main Results:
- The developed method demonstrated an unbiased and efficient DNA selection and amplification strategy.
- Successfully applied to a complex transcriptome library (GIS-PET).
Conclusions:
- The novel bacterial selection followed by MDA offers an improved approach for DNA library amplification.
- This method enhances efficiency and reduces bias in genomic and cDNA sequencing analyses.
