IMPROVED METHOD OF DNA REUSING BASED ON THE MULTIPLE DISPLACEMENT AMPLIFICATION AND STREPTAVIDIN MODIFIED MAGNETIC BEADS IMMOBILIZATION TECHNOLOGY

摘要

The fundament of almost every molecular biology assay is the availability of the adequate quality and quantity of genomic DNA. In many situations there may not be sufficient DNA collected from patient or population cohorts to meet the requirements of highly development assay technology. In this instance, Whole Genome Amplification (WGA) methods are used to generate the large amounts of DNA that are required for genetic testing. Many methods of WGA have been reported, such as Primer extension preamplification (PEP), degenerate-oligonucleotideprimed PCR (DOP) and inter-Alu-PCR. In recent years, many papers reported the isothermal multiple displacement amplification (MDA) technology, which could yield about 20-30 /ig product from as few as 1-10 copies of human genomic DNA. In this study, we developed a new method of DNA amplification and reusing based on the improved MDA and streptavidin modified magnetic beads immobilization technology. With this method, it is just required one sixth to one third amplification time compared with the traditional MDA method and the amplification can be carried out directly from biological samples including crude whole blood and tissue culture cells without the traditional DNA purification process.