[Evaluation of the fidelity of multiple displacement amplification from small number of cells].
OBJECTIVE
To evaluate the fidelity of multiple displacement amplification (MDA) from small number of cells (1-10 cells) by 10K 2.0 SNP mapping array.


METHODS
A 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.


RESULTS
The 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).


CONCLUSIONS
MDA 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.
Ling
Jiawei
J
Department of Reproductive Medicine, the First Affiliated Hospital of SUN Yat-sen University, Guangzhou, Guangdong, 510080 P.R. China. lingjiawei@yahoo.com
Fang
Cong
C
Xu
Yanwen
Y
Zhuang
Guanglun
G
Cao
Baoqiang
B
D002460:Cell Line; D002477:Cells / Q000166:cytology*; D004247:DNA / Q000235:genetics; D006801:Humans; D019656:Loss of Heterozygosity; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D013698:Templates, Genetic
D004247:DNA