| 主权项 |
1. A method of amplifying the whole genome of a single cell comprising
(a) providing genomic DNA from the single cell in single stranded form in a reaction vessel; (b) adding primers having a common sequence, a variable sequence and a fixed sequence to the reaction vessel to produce a reaction mixture, wherein the common sequence includes G, T and A but not C, (c) subjecting the reaction mixture to a low temperature at which annealing of the primers to the single stranded genomic DNA takes place, (d) adding at least one of a DNA polymerase having strand displacement activity or 5′ to 3′ exonuclease activity to the reaction mixture and subjecting the reaction mixture to a temperature at which DNA amplification takes place to produce single or double stranded DNA, (e) subjecting the reaction mixture to a temperature to produce single stranded amplicons, (f) optionally subjecting the reaction mixture to a temperature to anneal free primer to the 3′ end of amplicons thereby maintaining a linear structure to prevent the formation of chimeras, (g) repeating steps (c) to (f) to produce amplicons of the genomic DNA, and (h) analyzing the genomic DNA for congenital disorders or known phenotypical consequences including whole chromosome level abnormalities, chromosome translocations, aneuploidy, deletion or duplication of part or the whole chromosome, beta-thalassaemia Down's syndrome cystic fibrosis, sickle cell disease, tay-sachs disease, fragile X syndrome, spinal muscular atrophy, haemoglobinopathies, alpha-thalassemia, X-linked disorders (disorders determined by genes on the X chromosome), spina bifida, anencephaly, congenital heart defects, obesity, diabetes, cancer, fetal gender, fetal RHD, fetal HLA haplotype, paternally derived mutations, or chromosomal aneuploidy. |
