[Study on the amplification uniformity of whole genome in a single cell by degenerate oligonucleotide primed polymerase chain reaction].
OBJECTIVE
To investigate the uniformity of whole genome amplification from a single cell by degenerate oligonucleotide primed polymer ase chain reaction (DOP-PCR).


METHODS
Amplification of the whole genomic DNA from a single cell with XX, XY, XO, XXY, +13 or +21 was performed by DOP-PCR. Comparative genomic hybridization (CGH) of a single cell DOP-PCR product against the genomic DNA or a single cell DOP-PCR product from normal male as carried out respectively.


RESULTS
The average profile of fluorescence intensive ratio in CGH with the genomic DNA as the reference was greatly fluctuated and the standard deviation in about 30% haploid was beyond the normal limits. False positive hyperrepresentation was existed in X chromosome while trisomy 13 and 21 were failed to be detected. However, the distributions of the mean and the standard deviation of the ratio in the CGH with DOP-PCR product as the reference were acceptable. The copy number changes of chromosome X, Y, 13 and 21 were demonstrated.


CONCLUSIONS
Although the single cell DOP-PCR product is not completely uniform, it would be reliable to study the whole genome of a single cell by controlling the conditions of PCR and selecting the suitable control reference because the unequal amplification is unrandom.
Jin
F
F
Department of Obstetrics, Affiliated Obstetrics and Gynecology Hospital, Faculty of Medicine, Zhejiang University, Hangzhou 310006, China.
Huang
H
H
Ye
Y
Y
D005260:Female; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D019836:Preimplantation Diagnosis