The Establishiment And Evaluation Of The Methods For Single-Cell Whole Genome

The ultimate biological unit lies within a single cell. The secret trigger that signal mammal maturation, regeneration and genetic diseases all lie buried in a single cell that was originally part of the genetically clonal, multicellular organism. Many biological disciplines have taken aim to elucidate the causes of cellular differentiation at this level. Moreover, every single cell has its own unique microenvironment, there aren't two cells which are exactly the same. Beacuse of the heterogeneity of the tissues, it's necessary to analysis them at single-cell level, especially in the solid tumour. Recently, there are some researchers studied single cell using high-throughput analysis techniques with single cell isolating techniques and whole genome amplification methods.The techniques now used to isolate single cell are limited in cell suspensions. When isolating single cell from tissue like solid tomour, we have to digest the tissue into single cell. However, the location of the isolated cell is very important for same researchs. As to the case, we establish a method to isolate single complate nucleus from the frozen tissue section by laser microdissection which can solve this problem.There is little amount of DNA in a single cell. A human cell contains only about 6pg of genomic DNA. Whole genome amplification (WGA) offers means to overcome the above restrictions for single-cell genomic analyses. In this paper we used GenomePlex library method to amplified the DNA from single cell. Furthermore, we checked the efficiency,reproducibility,fedelity and the linear of the amplification product and we found it's suitable for downstream high-throughput analysis.In conclusion, we establish the methods to isolate single complate cell nucleus from the frozen tissue section and amplify the cell to get enough DNA for high-throughput analysis.