Full Text Available
Note: Clicking the button above will open the full text document at the original institutional repository in a new window.
Die identifikasie en gebruike van polimorfiese mikrosatellietvolgordes in genetiese kartering [Microfiche]
Thesis (MScAgric) -- Stellenbosch University, 1994.
Saved in:
| Main Author: | |
|---|---|
| Other Authors: | |
| Format: | Thesis |
| Language: | Afrikaans |
| Published: |
Stellenbosch : Stellenbosch University
2012
|
| Subjects: | |
| Tags: |
No Tags, Be the first to tag this record!
|
| _version_ | 1867613918465097729 |
|---|---|
| access_status_str | Open Access |
| author | Groenewald, I. M |
| author2 | Warnich, Louise |
| author_browse | Groenewald, I. M Warnich, Louise |
| author_facet | Warnich, Louise Groenewald, I. M |
| author_sort | Groenewald, I. M |
| collection | Thesis |
| dc_rights_str_mv | Stellenbosch University |
| description | Thesis (MScAgric) -- Stellenbosch University, 1994. |
| format | Thesis |
| id | oai:scholar.sun.ac.za:10019.1/54607 |
| institution | Stellenbosch University (South Africa) |
| language | Afrikaans |
| last_indexed | 2026-06-10T12:43:46.817Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository |
| publishDate | 2012 |
| publishDateRange | 2012 |
| publishDateSort | 2012 |
| publisher | Stellenbosch : Stellenbosch University |
| publisherStr | Stellenbosch : Stellenbosch University |
| record_format | dspace |
| source_str | SUNScholar — Stellenbosch University Repository |
| spelling | oai:scholar.sun.ac.za:10019.1/54607 Die identifikasie en gebruike van polimorfiese mikrosatellietvolgordes in genetiese kartering [Microfiche] Groenewald, I. M Warnich, Louise Retief, A. E. Stellenbosch University. Faculty of AgriSciences. Dept. of Genetics and Institute of Plant Biotechnology. Gene mapping Genetic polymorphisms Dissertations -- Genetics Thesis (MScAgric) -- Stellenbosch University, 1994. In this study 19 unique DNA clones, which have already been published and assigned to specific chromosomal regions and genetic maps, were examined for the presence of nine different microsatellite sequences. The aim of this study was to improve the usefulness of these markers, that show RFLP’s in the human genome, in linkage studies. Synthesised microsatellite sequences were radio-actively labelled for hybridization to filters containing restriction digests of the 19 clones. Eight clones showed hybridization with a (CA)12-oligonucleotide probe and one each with a (GA)10- and (AAAT)5-oligonucleotide probe. Three clones viz. E141, E24.1 and M8 that hybridized with the (CA)12-oligonucleotide probe and M8 that also hybridized with the (AAAT)5-oligonucleotide probe, were selected for further analysis. E141(CA)n, M8(CA)n and M8(AAAT)n, cloned in bacteriophages, were recloned in plasmid vectors for DNA sequencing with either degenerate primers (in the case of clones with CA-repeats) or bacteriophage M13 primers (in the case of M8(AAAT)n). Sequencing of the clone E24.1 (originally cloned in a plasmid) was also performed by means of degenerate primers. E141, M8 and E24.1 showed perfect CA-repeats of 15, 10 and 7 repeats respectively. M8 also contained a perfect AAAT-repeat that occurred twice. Unique primers were designed and synthesised for polymerase chain reaction (PCR) amplification of the repeat-containing regions of E24.1 and E141. The E24.1(CA)7 repeat was not polymorphic after analyses of 20 unrelated CEPH individuals. These results confirm the finding of Weber (1990b) that ten or less repeats are not likely to be polymorphic in the genome. For this reason further studies were not undertaken with M8(CA)10 and M8(AAAT)2. Analysis of the CA-repeat containing region of the E141(CA)n marker showed six different alleles in the CEPH families, inherited in Mendelian fashion. The observed heterozygosity of E141(CA) was calculated as 0,66 and the polymorphic information content (PIC) as 0,63. The original E141 (Eco RI) RFLP studies showed an observed heterozygosity of 0,53 and a PIC of 0,37 (Warnich et al., 1990e). Ten large CEPH families were used for PCR amplification of the repeat-containing region. Linkage studies were performed by means of the LINKAGE computer programmes and marker data from the CEPH database (version 3). The localization of the E141(CA) marker was determined on chromosome 5q33-q34 by two-point and multi-point linkage analyses with other markers on chromosome 5. The use of the highly polymorphic CA-microsatellite repeat has improved the usefulness of the locus D5S99 (E141) in linkage studies. Not only has the PIC and heterozygosity of this marker been improved, but an easy and fast method of genotyping by means of the PCR based assay, has been established. Masters 2012-08-27T11:36:38Z 2012-08-27T11:36:38Z 1994 Thesis http://hdl.handle.net/10019.1/54607 af Stellenbosch University 97 pages application/pdf Stellenbosch : Stellenbosch University |
| spellingShingle | Gene mapping Genetic polymorphisms Dissertations -- Genetics Groenewald, I. M Die identifikasie en gebruike van polimorfiese mikrosatellietvolgordes in genetiese kartering [Microfiche] |
| title | Die identifikasie en gebruike van polimorfiese mikrosatellietvolgordes in genetiese kartering [Microfiche] |
| title_full | Die identifikasie en gebruike van polimorfiese mikrosatellietvolgordes in genetiese kartering [Microfiche] |
| title_fullStr | Die identifikasie en gebruike van polimorfiese mikrosatellietvolgordes in genetiese kartering [Microfiche] |
| title_full_unstemmed | Die identifikasie en gebruike van polimorfiese mikrosatellietvolgordes in genetiese kartering [Microfiche] |
| title_short | Die identifikasie en gebruike van polimorfiese mikrosatellietvolgordes in genetiese kartering [Microfiche] |
| title_sort | die identifikasie en gebruike van polimorfiese mikrosatellietvolgordes in genetiese kartering microfiche |
| topic | Gene mapping Genetic polymorphisms Dissertations -- Genetics |
| url | http://hdl.handle.net/10019.1/54607 |
| work_keys_str_mv | AT groenewaldim dieidentifikasieengebruikevanpolimorfiesemikrosatellietvolgordesingenetiesekarteringmicrofiche |