Open AccessThe use of multiple displacement amplification to amplify complex DNA libraries
Author(s) -
Melissa J. Fullwood,
Jack J. S. Tan,
Patrick W. P. Ng,
Kuo Ping Chiu,
Jun Liu,
Chia Lin Wei,
Yijun Ruan
Publication year - 2008
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/gkn074
Subject(s) - biology , multiple displacement amplification , genomic library , insert (composites) , complementary dna , dna , computational biology , library , genomic dna , cdna library , selection (genetic algorithm) , ligation , genetics , microbiology and biotechnology , polymerase chain reaction , gene , dna extraction , base sequence , computer science , 16s ribosomal rna , artificial intelligence , mechanical engineering , engineering
Complex libraries for genomic DNA and cDNA sequencing analyses are typically amplified using bacterial propagation. To reduce biases, large numbers of colonies are plated and scraped from solid-surface agar. This process is time consuming, tedious and limits scaling up. At the same time, multiple displacement amplification (MDA) has been recently developed as a method for in vitro ampli- fication of DNA. However, MDA has no selection function for the removal of ligation multimers. We developed a novel method of briefly introducing ligation reactions into bacteria to select single insert DNA clones followed by MDA to amplify. We applied these methods to a Gene Identification Signatures with Paired-End diTags (GIS-PET) library, which is a complex transcriptome library created by pairing short tags from the 5' and 3' ends of cDNA fragments together, and demonstrated that this selection and amplification strategy is unbiased and efficient.
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