Development And Analysis Algorithm Research Of A New Generation A-CGH Arrays

With the implementation of the two-child policy in 2016 and the approaching of population aging in recent years,the detection of complex diseases,such as prenatal diagnosis and cancer are becoming more and more urgent.Genomic variations such as single nucleotide polymorphisms have been shown to cause various diseases,but most of complex diseases such as cancer remain unexplained.The discovery of gene copy number variations provide a new angle for us to look at variations in the genomes.Scientists have found that a large number of CNVs are so closely related to diseases that CNVs detection is becoming more and more important.The completion of human genome project in early 21st century greatly promoted the development of a new generation of gene detection methods such as gene chips.The whole genome chip is a full spectrum detection method that scan the whole genome to detect a large number of gene mutation simultaneously.The new generation chip of array-comparative genomic hybridization has the feature of high throughput,automation,high sensitivity,ease of operation and parallelism.Therefor we carried out the study.In this study,we developed the new generation of whole genome a-CGH arrays and performed data analysis.Major work include probe preparation,probe fix and later hybridization and data analysis.The main research contents are as follows:(1)Amplification of the new generation of a-CGH arrays probes with the isothermal amplification technique of Bst DNA polymerase large fragment.We had successfully produced 70ml concentrated Bst polymerase enzyme in twenty batch of cell culture,induced expression,purification and concentration.(2)Processing of a-CGH arrays probe:23033 groups of probe sets comprising of a total of over 17 millions specific oligo probe sequence were processed with isothermal amplification,a series of biological and chemical treatment.The probe set concentration be measured and adjusted equally to suitable for fixing on the substrate.(3)Hybridization testing of the new generation of a-CGH arrays:the sample DNA amplified Isothermally then labled with Cy3 and Cy5 fluorescent dyes,and then hybridized to the new generation of a-CGH arrays and scanned.(4)Analysis of the data of the new generation of a-CGH arrays:Location file created by setting up chip parameter to convert each probe spot on the scanned a-CGH arrays into data,then an algorithm was develop to analyze the data to get the copy number variation which can be visualized and data mined.This research has great significance in the development of major disease detection in china.Through the improvement of the experimental program,a new generation of whole genome a-CGH arrays was successfully developed.The a-CGH arrays can detect the copy number variation of the whole set of chromosomes and make use of the algorithm to get the visual map of the copy number variation of chromosome specific location.Formed a set of experimental and analysis process.