Identifying the sex of human preimplantation embryos in x-linked disease: Amplification efficiency of a Y-specific alphoid repeat from single blastomeres with two lysis protocols
| ISSN: |
1573-7330
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| Keywords: |
polymerase chain reaction ; single cell ; DNA amplification ; human preimplantation embryo ; preimplantation diagnosis ; sexing ; alphoid repeat
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| Source: |
Springer Online Journal Archives 1860-2000
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| Topics: |
Medicine
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| Notes: |
Abstract Introduction: Preimplantation diagnosis involves detecting genetic defects in one or two blastomeres biopsied from cleavage stage embryos followingin vitro fertilization (IVF). For X-linked recessive disease, identification of the sex of embryos allows transfer of only unaffected females. To examine how critical the preparation of the single blastomere is for amplification of a Y chromosome specific repeat sequence using the polymerase chain reaction (PCR), the incidence of amplification failure has been examined following two lysis protocols. Materials and Methods: Amplification of a Y alphoid repeat sequence from single blastomeres disaggregated from cleavage stage embryos was examined after either (1) lysis in distilled water and freeze-thawing twice or (2) a two-step lysis protocol involving an initial treatment in potassium hydroxide and dithiothreitol. Some of the embryos had been previously sexed by cleavage-stage biopsy and fluorescent in situ hybridization with X- and Y-specific probes. Results: Amplification failure occurred in 6 of 50 (12%) and 4 of 60 (7%) single blastomeres from male embryos following lysis in distilled water or using the two-step protocol, respectively. Conversely, amplification from contaminating DNA occurred in 5 of 63 (8%) single blastomeres from female embryos and 6 of 94 (6%) of control medium blanks. Conclusions: The incidence of amplification failure was improved but not eliminated using the two-step lysis protocol. At least two cells, therefore, would be necessary for accurate identification of males by amplification of Y-specific repeat sequences alone. Nevertheless, this protocol for preparing cleavage-stage blastomeres is likely to give more consistent amplification of any unique or repeat sequences.
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| Type of Medium: |
Electronic Resource
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| URL: |
| _version_ | 1798296904054865920 |
|---|---|
| autor | Kontogianni, E. H. Griffin, D. K. Handyside, A. H. |
| autorsonst | Kontogianni, E. H. Griffin, D. K. Handyside, A. H. |
| book_url | http://dx.doi.org/10.1007/BF02072533 |
| datenlieferant | nat_lic_papers |
| hauptsatz | hsatz_simple |
| identnr | NLM197487440 |
| issn | 1573-7330 |
| journal_name | Journal of assisted reproduction and genetics |
| materialart | 1 |
| notes | Abstract Introduction: Preimplantation diagnosis involves detecting genetic defects in one or two blastomeres biopsied from cleavage stage embryos followingin vitro fertilization (IVF). For X-linked recessive disease, identification of the sex of embryos allows transfer of only unaffected females. To examine how critical the preparation of the single blastomere is for amplification of a Y chromosome specific repeat sequence using the polymerase chain reaction (PCR), the incidence of amplification failure has been examined following two lysis protocols. Materials and Methods: Amplification of a Y alphoid repeat sequence from single blastomeres disaggregated from cleavage stage embryos was examined after either (1) lysis in distilled water and freeze-thawing twice or (2) a two-step lysis protocol involving an initial treatment in potassium hydroxide and dithiothreitol. Some of the embryos had been previously sexed by cleavage-stage biopsy and fluorescent in situ hybridization with X- and Y-specific probes. Results: Amplification failure occurred in 6 of 50 (12%) and 4 of 60 (7%) single blastomeres from male embryos following lysis in distilled water or using the two-step protocol, respectively. Conversely, amplification from contaminating DNA occurred in 5 of 63 (8%) single blastomeres from female embryos and 6 of 94 (6%) of control medium blanks. Conclusions: The incidence of amplification failure was improved but not eliminated using the two-step lysis protocol. At least two cells, therefore, would be necessary for accurate identification of males by amplification of Y-specific repeat sequences alone. Nevertheless, this protocol for preparing cleavage-stage blastomeres is likely to give more consistent amplification of any unique or repeat sequences. |
| package_name | Springer |
| publikationsjahr_anzeige | 1996 |
| publikationsjahr_facette | 1996 |
| publikationsjahr_intervall | 8004:1995-1999 |
| publikationsjahr_sort | 1996 |
| publisher | Springer |
| reference | 13 (1996), S. 125-132 |
| schlagwort | polymerase chain reaction single cell DNA amplification human preimplantation embryo preimplantation diagnosis sexing alphoid repeat |
| search_space | articles |
| shingle_author_1 | Kontogianni, E. H. Griffin, D. K. Handyside, A. H. |
| shingle_author_2 | Kontogianni, E. H. Griffin, D. K. Handyside, A. H. |
| shingle_author_3 | Kontogianni, E. H. Griffin, D. K. Handyside, A. H. |
| shingle_author_4 | Kontogianni, E. H. Griffin, D. K. Handyside, A. H. |
| shingle_catch_all_1 | Kontogianni, E. H. Griffin, D. K. Handyside, A. H. Identifying the sex of human preimplantation embryos in x-linked disease: Amplification efficiency of a Y-specific alphoid repeat from single blastomeres with two lysis protocols polymerase chain reaction single cell DNA amplification human preimplantation embryo preimplantation diagnosis sexing alphoid repeat polymerase chain reaction single cell DNA amplification human preimplantation embryo preimplantation diagnosis sexing alphoid repeat Abstract Introduction: Preimplantation diagnosis involves detecting genetic defects in one or two blastomeres biopsied from cleavage stage embryos followingin vitro fertilization (IVF). For X-linked recessive disease, identification of the sex of embryos allows transfer of only unaffected females. To examine how critical the preparation of the single blastomere is for amplification of a Y chromosome specific repeat sequence using the polymerase chain reaction (PCR), the incidence of amplification failure has been examined following two lysis protocols. Materials and Methods: Amplification of a Y alphoid repeat sequence from single blastomeres disaggregated from cleavage stage embryos was examined after either (1) lysis in distilled water and freeze-thawing twice or (2) a two-step lysis protocol involving an initial treatment in potassium hydroxide and dithiothreitol. Some of the embryos had been previously sexed by cleavage-stage biopsy and fluorescent in situ hybridization with X- and Y-specific probes. Results: Amplification failure occurred in 6 of 50 (12%) and 4 of 60 (7%) single blastomeres from male embryos following lysis in distilled water or using the two-step protocol, respectively. Conversely, amplification from contaminating DNA occurred in 5 of 63 (8%) single blastomeres from female embryos and 6 of 94 (6%) of control medium blanks. Conclusions: The incidence of amplification failure was improved but not eliminated using the two-step lysis protocol. At least two cells, therefore, would be necessary for accurate identification of males by amplification of Y-specific repeat sequences alone. Nevertheless, this protocol for preparing cleavage-stage blastomeres is likely to give more consistent amplification of any unique or repeat sequences. 1573-7330 15737330 Springer |
| shingle_catch_all_2 | Kontogianni, E. H. Griffin, D. K. Handyside, A. H. Identifying the sex of human preimplantation embryos in x-linked disease: Amplification efficiency of a Y-specific alphoid repeat from single blastomeres with two lysis protocols polymerase chain reaction single cell DNA amplification human preimplantation embryo preimplantation diagnosis sexing alphoid repeat polymerase chain reaction single cell DNA amplification human preimplantation embryo preimplantation diagnosis sexing alphoid repeat Abstract Introduction: Preimplantation diagnosis involves detecting genetic defects in one or two blastomeres biopsied from cleavage stage embryos followingin vitro fertilization (IVF). For X-linked recessive disease, identification of the sex of embryos allows transfer of only unaffected females. To examine how critical the preparation of the single blastomere is for amplification of a Y chromosome specific repeat sequence using the polymerase chain reaction (PCR), the incidence of amplification failure has been examined following two lysis protocols. Materials and Methods: Amplification of a Y alphoid repeat sequence from single blastomeres disaggregated from cleavage stage embryos was examined after either (1) lysis in distilled water and freeze-thawing twice or (2) a two-step lysis protocol involving an initial treatment in potassium hydroxide and dithiothreitol. Some of the embryos had been previously sexed by cleavage-stage biopsy and fluorescent in situ hybridization with X- and Y-specific probes. Results: Amplification failure occurred in 6 of 50 (12%) and 4 of 60 (7%) single blastomeres from male embryos following lysis in distilled water or using the two-step protocol, respectively. Conversely, amplification from contaminating DNA occurred in 5 of 63 (8%) single blastomeres from female embryos and 6 of 94 (6%) of control medium blanks. Conclusions: The incidence of amplification failure was improved but not eliminated using the two-step lysis protocol. At least two cells, therefore, would be necessary for accurate identification of males by amplification of Y-specific repeat sequences alone. Nevertheless, this protocol for preparing cleavage-stage blastomeres is likely to give more consistent amplification of any unique or repeat sequences. 1573-7330 15737330 Springer |
| shingle_catch_all_3 | Kontogianni, E. H. Griffin, D. K. Handyside, A. H. Identifying the sex of human preimplantation embryos in x-linked disease: Amplification efficiency of a Y-specific alphoid repeat from single blastomeres with two lysis protocols polymerase chain reaction single cell DNA amplification human preimplantation embryo preimplantation diagnosis sexing alphoid repeat polymerase chain reaction single cell DNA amplification human preimplantation embryo preimplantation diagnosis sexing alphoid repeat Abstract Introduction: Preimplantation diagnosis involves detecting genetic defects in one or two blastomeres biopsied from cleavage stage embryos followingin vitro fertilization (IVF). For X-linked recessive disease, identification of the sex of embryos allows transfer of only unaffected females. To examine how critical the preparation of the single blastomere is for amplification of a Y chromosome specific repeat sequence using the polymerase chain reaction (PCR), the incidence of amplification failure has been examined following two lysis protocols. Materials and Methods: Amplification of a Y alphoid repeat sequence from single blastomeres disaggregated from cleavage stage embryos was examined after either (1) lysis in distilled water and freeze-thawing twice or (2) a two-step lysis protocol involving an initial treatment in potassium hydroxide and dithiothreitol. Some of the embryos had been previously sexed by cleavage-stage biopsy and fluorescent in situ hybridization with X- and Y-specific probes. Results: Amplification failure occurred in 6 of 50 (12%) and 4 of 60 (7%) single blastomeres from male embryos following lysis in distilled water or using the two-step protocol, respectively. Conversely, amplification from contaminating DNA occurred in 5 of 63 (8%) single blastomeres from female embryos and 6 of 94 (6%) of control medium blanks. Conclusions: The incidence of amplification failure was improved but not eliminated using the two-step lysis protocol. At least two cells, therefore, would be necessary for accurate identification of males by amplification of Y-specific repeat sequences alone. Nevertheless, this protocol for preparing cleavage-stage blastomeres is likely to give more consistent amplification of any unique or repeat sequences. 1573-7330 15737330 Springer |
| shingle_catch_all_4 | Kontogianni, E. H. Griffin, D. K. Handyside, A. H. Identifying the sex of human preimplantation embryos in x-linked disease: Amplification efficiency of a Y-specific alphoid repeat from single blastomeres with two lysis protocols polymerase chain reaction single cell DNA amplification human preimplantation embryo preimplantation diagnosis sexing alphoid repeat polymerase chain reaction single cell DNA amplification human preimplantation embryo preimplantation diagnosis sexing alphoid repeat Abstract Introduction: Preimplantation diagnosis involves detecting genetic defects in one or two blastomeres biopsied from cleavage stage embryos followingin vitro fertilization (IVF). For X-linked recessive disease, identification of the sex of embryos allows transfer of only unaffected females. To examine how critical the preparation of the single blastomere is for amplification of a Y chromosome specific repeat sequence using the polymerase chain reaction (PCR), the incidence of amplification failure has been examined following two lysis protocols. Materials and Methods: Amplification of a Y alphoid repeat sequence from single blastomeres disaggregated from cleavage stage embryos was examined after either (1) lysis in distilled water and freeze-thawing twice or (2) a two-step lysis protocol involving an initial treatment in potassium hydroxide and dithiothreitol. Some of the embryos had been previously sexed by cleavage-stage biopsy and fluorescent in situ hybridization with X- and Y-specific probes. Results: Amplification failure occurred in 6 of 50 (12%) and 4 of 60 (7%) single blastomeres from male embryos following lysis in distilled water or using the two-step protocol, respectively. Conversely, amplification from contaminating DNA occurred in 5 of 63 (8%) single blastomeres from female embryos and 6 of 94 (6%) of control medium blanks. Conclusions: The incidence of amplification failure was improved but not eliminated using the two-step lysis protocol. At least two cells, therefore, would be necessary for accurate identification of males by amplification of Y-specific repeat sequences alone. Nevertheless, this protocol for preparing cleavage-stage blastomeres is likely to give more consistent amplification of any unique or repeat sequences. 1573-7330 15737330 Springer |
| shingle_title_1 | Identifying the sex of human preimplantation embryos in x-linked disease: Amplification efficiency of a Y-specific alphoid repeat from single blastomeres with two lysis protocols |
| shingle_title_2 | Identifying the sex of human preimplantation embryos in x-linked disease: Amplification efficiency of a Y-specific alphoid repeat from single blastomeres with two lysis protocols |
| shingle_title_3 | Identifying the sex of human preimplantation embryos in x-linked disease: Amplification efficiency of a Y-specific alphoid repeat from single blastomeres with two lysis protocols |
| shingle_title_4 | Identifying the sex of human preimplantation embryos in x-linked disease: Amplification efficiency of a Y-specific alphoid repeat from single blastomeres with two lysis protocols |
| sigel_instance_filter | dkfz geomar wilbert ipn albert fhp |
| source_archive | Springer Online Journal Archives 1860-2000 |
| timestamp | 2024-05-06T09:59:29.679Z |
| titel | Identifying the sex of human preimplantation embryos in x-linked disease: Amplification efficiency of a Y-specific alphoid repeat from single blastomeres with two lysis protocols |
| titel_suche | Identifying the sex of human preimplantation embryos in x-linked disease: Amplification efficiency of a Y-specific alphoid repeat from single blastomeres with two lysis protocols |
| topic | WW-YZ |
| uid | nat_lic_papers_NLM197487440 |