- Author:
Jiawei LING
1
;
Cong FANG
;
Yanwen XU
;
Guanglun ZHUANG
;
Baoqiang CAO
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line; Cells; cytology; DNA; genetics; Humans; Loss of Heterozygosity; Nucleic Acid Amplification Techniques; methods; Polymorphism, Single Nucleotide; Templates, Genetic
- From: Chinese Journal of Medical Genetics 2010;27(1):42-46
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo evaluate the fidelity of multiple displacement amplification (MDA) from small number of cells (1-10 cells) by 10K 2.0 SNP mapping array.
METHODSA fibroblast cell line (Tri-18; GM02732, 47, XY, +18) was used as the template, and 6 groups were set up in the study. Groups A and B were positive and negative control, respectively; groups C-F were experimental groups involving the MDA products from 1, 2, 5 and 10 cells respectively. In combination of single nucleotide polymorphism (SNP) array, the product of each group was assessed based on the genome coverage, loss of heterozygosity (LOH) rate and allele dropout (ADO) rate.
RESULTSThe nonspecific product of negative control presented an average call rate of 3.2%. The genome coverage of the MDA product increased from 86.4% to 96.4% with the increasing number of template from 1 to 10 cells, while the LOH rate and ADO rate decreased significantly (P<0.05).
CONCLUSIONMDA is a highly efficient and reliable method for whole genome amplification. The fidelity of MDA will be improved significantly with the increasing number of template cells. 10K 2.0 SNP mapping array is a quick, accurate and comprehensive method to evaluate the fidelity of amplified DNA products, but the ADO SNPs should be distinguished from those of preferential amplification from the LOH loci to avoid errors.