PGD requires the use of assisted reproduction technology (ART) and it has already been used since the beginning of the 90s, initially applied for monogenic diseases [5] and shortly after for chromosomal rearrangements [6]. all single-cell samples are depicted: (A) dup(7)(p14.3p21.3) G0/G1-phase cells. 1755-8166-7-46-S2.png (159K) GUID:?D952A0ED-1423-4EC2-BF0F-CB978A93A89A Additional file 3 aCGH profiles of the derivative chromosomes for all the single cells analyzed. aCGH plots of the chromosomes of interest with fluorescence intensity log2 ratios on the X-axis and chromosomal position on the Y-axis. Plots for all single-cell samples are depicted: (B) dup(7)(p14.3p21.3) S-phase cells. 1755-8166-7-46-S3.png (215K) GUID:?354748F9-64C5-4189-83D5-376CA7E3240B Additional file 4 aCGH profiles of the derivative chromosomes for all the single cells analyzed. aCGH plots of the chromosomes of interest with fluorescence intensity log2 ratios on the X-axis and chromosomal position on the Y-axis. Plots for all single-cell samples are depicted: (C) der(18)t(9;18)(p21.3;p11.3) G0/G1-phase cells. 1755-8166-7-46-S4.png (263K) GUID:?6274E80C-E23F-4992-829B-F2618898DAD1 Additional file 5 aCGH profiles of the derivative chromosomes for all the single cells analyzed. aCGH plots of the chromosomes of interest with fluorescence intensity log2 ratios on the X-axis and Rabbit Polyclonal to DGKZ chromosomal position on the Y-axis. Plots for all single-cell samples are depicted: (D) der(18)t(9;18)(p21.3;p11.3) S-phase cells. 1755-8166-7-46-S5.png (353K) GUID:?2459D218-0CBB-4E11-A7B7-B9225EDBD7AB Additional file 6 aCGH profiles of the derivative chromosomes for all the single cells analyzed. aCGH plots of the chromosomes of interest with fluorescence intensity log2 ratios on the X-axis and chromosomal position on the Y-axis. Plots for all single-cell samples are depicted: (E) der(20)t(18;20)(p11.22;p13) G0/G1-phase cells. 1755-8166-7-46-S6.png (214K) GUID:?7BF64A82-9D20-404D-878A-4858986B37A9 Additional file 7 aCGH profiles of the derivative chromosomes for all the single cells analyzed. aCGH plots of the chromosomes of interest with fluorescence intensity log2 ratios on the X-axis and chromosomal position on the Y-axis. Plots for all single-cell samples are depicted: (F) der(20)t(18;20)(p11.22;p13) S-phase cells. 1755-8166-7-46-S7.png (208K) GUID:?1033ADA5-81A9-43D3-A44B-9F82AEB1E09E Additional file 8 Estimation of false positive detection rate of the derivative chromosomes in Deltasonamide 2 control single S- and G0/G1-phase cells of three cell lines. Number of probes falsely indicating the presence Deltasonamide 2 of an imbalance in the regions of interest according to the BAC array log2 intensity ratios in S-phase single cells of cell lines not carrying an aberration involving this region. In brackets the number of probes in the regions of interest. *The log2 intensity ratios were indicative of a deletion in the control sample, while there is a duplication in the same region in the cell line of interest. 1755-8166-7-46-S8.xlsx (9.7K) GUID:?B5EE69CF-96F0-4FC3-85EC-CDE254B3BC95 Additional file 9 Cell sorting procedure. Representative plot illustrating the cell sorting procedure by FACS with relative DNA content on the X-axis and the cell count on the Y-axis. The marked windows correspond to the fractions which were collected for each cell subpopulation (G0/G1-, S- and G2/M-phase). 1755-8166-7-46-S9.png (190K) GUID:?577A2C36-2BA8-48CB-B267-EA2356CBC6CE Abstract Background Carriers of balanced translocations are at high risk for unbalanced gametes which can result in recurrent miscarriages or birth defects. Preimplantation genetic diagnosis (PGD) is often offered to select balanced embryos. This selection is currently mainly performed by array CGH on blastomeres. Current methodology does not take into account the phase of the cell cycle, despite the variable copy number status of different genomic regions in S phase. Results Cell lines derived from 3 patients with different chromosomal imbalances were used to evaluate the accuracy of single cell array CGH. The different cell cycle phases were sorted by flow cytometry and 10 single cells were picked per cell line per cell cycle phase, whole genome amplified and Deltasonamide 2 analyzed by BAC arrays, the most commonly used platform for PGD purposes. In contrast to G phase, where the imbalances were efficiently identified, less than half of the probes in the regions of interest indicated the presence of the aberration in 17 S-phase cells, resulting in reduced accuracy. Conclusions The results demonstrate that the accuracy to detect segmental chromosomal imbalances is reduced in S-phase cells, which could be a source of misdiagnosis in PGD. Hence, the cell cycle Deltasonamide 2 phase of the analyzed cell is of great importance and should be taken into account during the analysis. This knowledge may guide future technological improvements. Background Up to 15% of the couples confront fertility problems and 1% of the couples attempting to conceive a child experience recurrent miscarriage (RM), defined as.