CLC number: R394.3
On-line Access:
Received: 2000-11-20
Revision Accepted: 2001-02-20
Crosschecked: 0000-00-00
Cited: 0
Clicked: 4549
JIN Fan, Matthews, C.D, Hussey, D.N. SINGLE CELL DEGENERATE OLIGONUCLEOTIDE PRIMER-PCR AND COMPARATIVE GENOMIC HYBRIDIZATION WITH MODIFIED CONTROL REFERENCE[J]. Journal of Zhejiang University Science A, 2001, 2(3): 318-321.
@article{title="SINGLE CELL DEGENERATE OLIGONUCLEOTIDE PRIMER-PCR AND COMPARATIVE GENOMIC HYBRIDIZATION WITH MODIFIED CONTROL REFERENCE",
author="JIN Fan, Matthews, C.D, Hussey, D.N",
journal="Journal of Zhejiang University Science A",
volume="2",
number="3",
pages="318-321",
year="2001",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.2001.0318"
}
%0 Journal Article
%T SINGLE CELL DEGENERATE OLIGONUCLEOTIDE PRIMER-PCR AND COMPARATIVE GENOMIC HYBRIDIZATION WITH MODIFIED CONTROL REFERENCE
%A JIN Fan
%A Matthews
%A C.D
%A Hussey
%A D.N
%J Journal of Zhejiang University SCIENCE A
%V 2
%N 3
%P 318-321
%@ 1869-1951
%D 2001
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2001.0318
TY - JOUR
T1 - SINGLE CELL DEGENERATE OLIGONUCLEOTIDE PRIMER-PCR AND COMPARATIVE GENOMIC HYBRIDIZATION WITH MODIFIED CONTROL REFERENCE
A1 - JIN Fan
A1 - Matthews
A1 - C.D
A1 - Hussey
A1 - D.N
J0 - Journal of Zhejiang University Science A
VL - 2
IS - 3
SP - 318
EP - 321
%@ 1869-1951
Y1 - 2001
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2001.0318
Abstract: For investigating the possibility of applying degenerate oligonucleotide primer PCR (DOP-PCR) and comparative genomic hybridization (CGH) technique to analyses of genomic genetics in a single cell, the whole genomic DNA of a single cell with XX, XY, XO, XXY, +13 or +21 was amplified by DOP-PCR. single cell DOP-PCR CGHs with conventional and modified control references, the genomic DNA and a single cell DOP-PCR product from normal male, were carried out respectively. The results showed that the average profile of the fluorescence intensity ratio in CGH with the genomic DNA as reference fluctuates much and that the standard deviation in about 30% haploid is beyond the normal limits. False positive hyper-representation was found to exist in X chromosome while trisomy 13 and 21 were not detected. However, the distributions of the mean and the standard deviation of the ratio in the CGH with DOP-PCR product as reference were quite acceptable. The copy number changes of chromosome X,Y,13 and 21 were revealed. Those results suggested that there is unrandom unequal amplification in a single cell DOP-PCR. Using a single DOP-PCR product as reference can decrease its influence on CGH. single cell DOP-PCR-CGH and its application in the genetic analyses of preimplantation embryo or fetal cell in maternal blood may be possible.
[1] Cheung V.G, Nelson S.F, 1996. Whole genome amplification using a degenerate oligonucleotide primer allows hundreds of genotypes to be performed on less than one nanogram of genomic DNA. Proc. Natl. Acad. Sci. USA, 93:14676-14679.
[2] Cui, K.H., Haan, E.A., Wang, L.J. et al., 1995. Optimal polymerase chain reaction amplification for preimplantation diagnosis in cystic fibrosis. BMJ, 311(7004):536-540.
[3] Harper, J.C., 1996. Preimplantation diagnosis of inherited disease by embryo biopsy: an update of the world figures. J Assist Reprod Genet, 13:90-95.
[4] Kallioniemi, A., Kallioniemi, O.P., Dudar D. et al., 1992. Comparative genomic hybridization for molecular cytogenetic analysis of solid tumors. Science, 258:818-821.
[5] Kallioniemi, A., Kallioniemi, O.P., Piper J. et al., 1994. Optimizing comparative genomic hybridization for analysis of DNA sequence copy number changes in solid tumors. Gene Chrom Cancer, 10:231-243.
[6] Kirchhoff, M, Gerdes T., Rose, H et al., 1998. Detection of chromosomal gains and losses in comparative genomic hybridization analysis based on standard reference intervals. Cytometry, 31:163-173.
[7] Kuukasjarvi, T., Tanner, M., Pennanen, S. et al., 1997. Optimizing DOP-PCR for univertsal amplification of small DNA samples in comparative genomic hybridization. Gene Chrom Cancer, 18:94-101.
[8] Sabtalo, J., Veiga, A., Calafell, J.M. et al., 1995. Evaluation of cytogenetic analysis for clinical preimplantation diagnosis. Fertil Steril, 64:44-50.
[9] Telenuis, H., Carter, N.P., Bebb, C.E. et al., 1992. Degenerate oligonucleotide-primed PCR: general amplification of target DNA by a single degenerate primer. Genomics, 13:718-725.
[10] Van-den-Veyver, I.B., 1998. Applied molecular genetic techniques for prenatal diagnosis. Curr Opin Obstet Gynecol, 10:97-103
[11] Verlinsky, Y., Munne S., Simpson J.L., 1997. Current status of preimplantation diagnosis. J Assist Reprod Genet, 14:72-75.
[12] Wells, D., Sherlock, J.K., Handyside, A.H. et al,1999. Detailed chromosomal and molecular genetic analysis of single cells by whole genome amplification and comparative genomic hybridisation. Nucleic Acids Res, 27:1214-1218.
[13] Wells, D., Delhanty, J.D, 2000. Comprehensive chromosomal analysis of human preimplantation embryos using whole genome amplification and single cell comparative genomic hybridization. Mol Hum Reprod, 6:1055-1062.
Open peer comments: Debate/Discuss/Question/Opinion
<1>