1. Preferential selection and transfer of euploid noncarrier embryos in preimplantation genetic diagnosis cycles for reciprocal translocations 
Li Wang, M.D., Ph.D., Jiandong Shen, M.D., David S. Cram, Ph.D., Minyue Ma, Ph.D., Hui Wang, M.D., Wenke Zhang, Ph.D., Junmei Fan, Ph.D., Zhiying Gao, M.D., Liwen Zhang, M.D., Zhifeng Li, Ph.D., Mengnan Xu, B.Sc., Don A. Leigh, Ph.D., Alan O. Trounson, Ph.D., Jiayin Liu, M.D., Ph.D., Yuanqing Yao, M.D., Ph.D. 
Fertility and Sterility 
Volume 108, Issue 4, Pages e4 (October 2017)
DOI: /j.fertnstert
Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Strategy for identification of noncarrier embryos in preimplantation genetic diagnosis (PGD) cycles for reciprocal translocations. (A) Mapping of reciprocal translocation breakpoints. Genomic DNA from the carrier parent is first subjected to mate pair sequencing to derive 5-kb fragments containing the breakpoint (BP) and nonbreakpoint (N) flanking sequences of the two involved chromosomes A and B. Alignment of the 5-kb fragments defines the minimal BP regions for segments ABʹ and BAʹ. Based on the known terminal sequences of the two minimal BP regions, a series of forward and reverse PCR primers (→) are designed progressively at 200-bp intervals toward the internally located BP. Nested PCR with all designed primer combinations is then performed to identify one pair that produces an amplicon containing the BP. If no amplicon is produced, a further set of primers () are designed progressively closer to the BP, and nested PCR is continued until an amplicon is produced. Further Sanger sequencing of these amplicons containing each of the BPs defines the exact BP junctions and flanking sequences. This sequence information is finally used to design and validate BP and N primers for PGD. (B) Clinical PGD with carrier testing. A blastocyst biopsy is performed and used for 24-chromosome testing to identify balanced/euploid embryos. The whole-genome amplification (WGA) products from these embryos are then further tested by PCR with the BP and N diagnostic primers to discriminate between carrier and noncarrier embryos. A noncarrier embryo is finally transferred, and the fetus from the ongoing pregnancy is tested by confirmatory prenatal diagnosis.
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3. Figure 2
Polymerase chain reaction (PCR) analysis to discriminate between carrier and noncarrier euploid embryos. Agarose-ethidium bromide gels showing the presence (+) or absence (−) of a PCR product for breakpoint (BP) or normal (N) sequences. B = blank PCR negative control; C = carrier; E = embryo; MW = molecular weight standards; NC = noncarrier; P1 = carrier DNA positive control; P2 = noncarrier DNA positive control.
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4. Figure 3
Summary of preimplantation genetic diagnosis (PGD) results for a 46,XY,t(7;14)(q22;q24.3) patient. (A) Translocation breakpoint identification. (B) High-resolution embryo analysis of translocation chromosomes. (C) Discrimination of carrier from noncarrier embryos. B = blank PCR negative control; C = carrier; E = embryo; MW = molecular weight standards; NC = noncarrier; P1 = carrier DNA positive control; P2 = noncarrier DNA positive control. (D) Family karyotypes.
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5. Supplemental Figure 1
24-chromosome profiles of balanced euploid embryos. (A) Array-based comparative genomic hybridization (aCGH) analysis of a blastomere biopsy sample. (B) Next-generation sequencing (NGS) analysis of a trophectoderm biopsy sample.
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6. Supplemental Figure 2
Karyotypes of parents and children born after preimplantation genetic diagnosis (PGD).
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7. Supplemental Figure 3
Identification of balanced translocation breakpoints. The DNA sequences juxtaposed between the arrowed lines (unknown origin) were presumably incorporated during the formation of the balanced translocation.
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8. Supplemental Figure 4
Analysis of allelic dropout (ADO) rates of breakpoint and nonbreakpoint primers for polymerase chain reaction (PCR) of 46,XY,t(7;14)(q22;q24.3) lymphocyte DNA (five cells). B = blank PCR negative control; P = DNA positive control.
Fertility and Sterility  , e4DOI: ( /j.fertnstert )
Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions