Application of multiple displacement amplification in detection of five events of genetically modified organisms

In this study, multiple displacement amplification (MDA) was used for transgene detection, and proved to effectively improve the sensitivity of most fluorescence polymerase chain (PCR). Five events of genetically modified plants were chosen for detection in different plant species by MDA and real-time PCRs. Individual primer/probe systems based on each transgene target were used. The extracted DNA of each sample was diluted and used as a template for MDA. All extracted DNA and diluted mdaDNA were detected by real-time PCR. The results were evaluated with respect to DNA quantity, DNA purity and difference of Ct values of the isogenetic DNA extract before and after MDA (D-values). The MDA amplifying effects were largely different among genes and plants. Organelle DNA has more copies than genomic DNA by MDA. Key words: Multiple displacement amplification, genetically modified organisms, detection, events.

[1]  De-xin Li,et al.  [Investigation of RNA viral genome amplification by multiple displacement amplification technique]. , 2013, Bing du xue bao = Chinese journal of virology.

[2]  Pavel A Pevzner,et al.  Genome of the pathogen Porphyromonas gingivalis recovered from a biofilm in a hospital sink using a high-throughput single-cell genomics platform , 2013, Genome research.

[3]  A. Romphruk,et al.  Development of a multiplex polymerase chain reaction-sequence-specific primer method for NKG2D and NKG2F single-nucleotide polymorphism typing using isothermal multiple displacement amplification products. , 2013, Tissue antigens.

[4]  Y. Chen,et al.  [Applying multiple displacement amplification to DNA typing in the pathological section]. , 2013, Fa yi xue za zhi.

[5]  Alexander Sczyrba,et al.  Decontamination of MDA Reagents for Single Cell Whole Genome Amplification , 2011, PloS one.

[6]  M. Fischer,et al.  DNA-based identification of spices: DNA isolation, whole genome amplification, and polymerase chain reaction. , 2011, Journal of agricultural and food chemistry.

[7]  W. Azmi,et al.  Multiple displacement amplification as a pre‐polymerase chain reaction (pre‐PCR) to detect ultra low population of Ralstonia solanacearum (Smith 1896) Yabuchi et al. (1996) , 2009, Letters in applied microbiology.

[8]  T. Hsiang,et al.  Whole genome amplification of the rust Puccinia striiformis f. sp. tritici from single spores. , 2009, Journal of microbiological methods.

[9]  Roger S Lasken,et al.  Genomic DNA amplification by the multiple displacement amplification (MDA) method. , 2009, Biochemical Society transactions.

[10]  C. Zhang,et al.  Preimplantation genetic diagnosis for Duchenne muscular dystrophy by multiple displacement amplification. , 2009, Fertility and sterility.

[11]  M. Lipinski,et al.  Multiple displacement amplification for complex mixtures of DNA fragments , 2008, BMC Genomics.

[12]  R. Kleespies,et al.  Sequencing of the large dsDNA genome of Oryctes rhinoceros nudivirus using multiple displacement amplification of nanogram amounts of virus DNA. , 2008, Journal of virological methods.

[13]  H. Broll,et al.  Generation of Reference Material by the Use of Multiple Displacement Amplification (MDA) for the Detection of Genetically Modified Organisms (GMOs) , 2008 .

[14]  T. Tengs,et al.  BMC Biotechnology BioMed Central Methodology article , 2008 .

[15]  Mark B Gerstein,et al.  Assessment of whole genome amplification-induced bias through high-throughput, massively parallel whole genome sequencing , 2006, BMC Genomics.

[16]  Tanja Dreo,et al.  Critical points of DNA quantification by real-time PCR--effects of DNA extraction method and sample matrix on quantification of genetically modified organisms. , 2006, BMC biotechnology.

[17]  J. Squire,et al.  The use of whole genome amplification in the study of human disease. , 2004, Progress in biophysics and molecular biology.

[18]  Roger S Lasken,et al.  Unbiased whole-genome amplification directly from clinical samples. , 2003, Genome research.

[19]  Daniel Pinkel,et al.  Whole genome analysis of genetic alterations in small DNA samples using hyperbranched strand displacement amplification and array-CGH. , 2003, Genome research.

[20]  S. Kingsmore,et al.  Comprehensive human genome amplification using multiple displacement amplification , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Jasper Becker,et al.  Joint Research Centre , 1982, Nature.