A Technique For Preimplantation Genetic Screening Based On Transposition

With the blooming of high-throughput sequencing technology,the Massively Parallel Sequencing?MPS?has been applied into medical realm much more widely nowadays.Such technology as non-invasive prenatal screening for Down Syndrome?Non-invasive Fetal Trisomy Test,NIFTY?and Pre-implantation Genetic Screening\ Diagnosis?PGS\PGD?are based on the next-generation sequencing.However,the conventional library construction methods for genome-sequencing cannot solve those “economic problems” like how to draw down the costs of manipulating tons of samples simultaneously and simplifying complicated procedures,even the labor resources should be involved into our consideration.What's worse,such methods cannot meet the demands of short periods detection.Therefore,to satisfy the growing needs for clinical detection,a set of low cost,high quality and rapid construction technology is eager to be developed.For recent years,the application of transposon has made great achievement since the in-depth research for transposition techniques,such as the Nextera^TM series of library preparation reagents from Illumina,have greatly reduced the redundant time for library constructing with their characteristics for being convenient and stable.This study is to develop a novel and rapid sequencing-library construction method exclusive for a new generation of semiconductor sequencing technology(Ion Proton^TM sequencing platform)possessed by Life Technology Corporation.Through designing the proprietary sequencing primer?Adapter sequence P1' and N' of the barcoded adapter?,following by the embedment and incubation of Tn5 transposase,to attack the desired DNA sample in order to accomplish the library preparation process.The total time for its whole procedure has been cut down to 1/5 of the original one,it also decreases the steps and apparatus during experiment and the time for DNA fragmentation.The samples of this study has involved following situations: DNA and single/multiple cell Whole Genome Amplification?WGA?products from cell line with chromosome deletion/duplication and Copy Number Variation?CNV?,and the WGA products of embryo samples.All of databases from original platform and clinical results are used to verify this study.Ultimately,by comparing the two approaches for library construction,both reach a 100% construction success rate and the coverage are 0.1X.Even for the CNV karyotype results and embryonic cell WGA products,they share 100% consistency.These outcomes manifest that such optimized library preparation method is feasible and can solve all problems mentioned above more efficiently by retaining the same performance.This technology is also conducive to promote new generation of mini single cell sequencing and microscale DNA fingerprinting technology.