Field collection,preservation and large scale DNA extraction procedures for cassava (Manihot esculenta Crantz.)
Some genetic studies using molecular methods such as diversity assessment or marker-assisted selection require collection of a large number of samples from fields located in the vicinity or in remote areas, followed by isolation of good quality DNA in a short time span. In the present study, differe...
| Main Authors: | , , , , |
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| Format: | Journal Article |
| Language: | Inglés |
| Published: |
2009
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10568/90807 |
| _version_ | 1855515992791187456 |
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| author | Bhattacharjee, Ranjana Ferguson, Morag E. Gedil, Melaku A Dumet, D. Ingelbrecht, I.L. |
| author_browse | Bhattacharjee, Ranjana Dumet, D. Ferguson, Morag E. Gedil, Melaku A Ingelbrecht, I.L. |
| author_facet | Bhattacharjee, Ranjana Ferguson, Morag E. Gedil, Melaku A Dumet, D. Ingelbrecht, I.L. |
| author_sort | Bhattacharjee, Ranjana |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Some genetic studies using molecular methods such as diversity assessment or marker-assisted selection require collection of a large number of samples from fields located in the vicinity or in remote areas, followed by isolation of good quality DNA in a short time span. In the present study, different tissue preservation methods were compared for subsequent DNA extraction using a modified CTAB method in two 96-well plates, following grinding of leaf tissues with a GenoGrinder 2000. We found that preservation of leaf tissues in NaCl-CTAB-azide buffer (as described in Rogstad, 1992) at 4°C is a better storage procedure than preservation at -20°C to obtain good quality DNA. Comparison of DNA extraction with or without use of phenol revealed that the quality of DNA was not drastically affected when non-phenol extraction protocol was used and did not affect PCR amplification. Thus, the recommended DNA extraction procedure allowed us to process 192 samples per day at a cost of $0.80 per sample, with an average yield of 1.8 g, suitable for both PCR and genotyping. |
| format | Journal Article |
| id | CGSpace90807 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2009 |
| publishDateRange | 2009 |
| publishDateSort | 2009 |
| record_format | dspace |
| spelling | CGSpace908072025-10-16T09:52:51Z Field collection,preservation and large scale DNA extraction procedures for cassava (Manihot esculenta Crantz.) Bhattacharjee, Ranjana Ferguson, Morag E. Gedil, Melaku A Dumet, D. Ingelbrecht, I.L. cassava nacl-ctab-azide solution phenol genogrinder Some genetic studies using molecular methods such as diversity assessment or marker-assisted selection require collection of a large number of samples from fields located in the vicinity or in remote areas, followed by isolation of good quality DNA in a short time span. In the present study, different tissue preservation methods were compared for subsequent DNA extraction using a modified CTAB method in two 96-well plates, following grinding of leaf tissues with a GenoGrinder 2000. We found that preservation of leaf tissues in NaCl-CTAB-azide buffer (as described in Rogstad, 1992) at 4°C is a better storage procedure than preservation at -20°C to obtain good quality DNA. Comparison of DNA extraction with or without use of phenol revealed that the quality of DNA was not drastically affected when non-phenol extraction protocol was used and did not affect PCR amplification. Thus, the recommended DNA extraction procedure allowed us to process 192 samples per day at a cost of $0.80 per sample, with an average yield of 1.8 g, suitable for both PCR and genotyping. 2009 2018-02-06T12:14:51Z 2018-02-06T12:14:51Z Journal Article https://hdl.handle.net/10568/90807 en Open Access Bhattacharjee, R., Ferguson, M., Gedil, M., Dumet, D. & Ingelbrecht, I. (2009). Field collection, preservation and large scale DNA extraction procedures for cassava (Manihot esculenta Crantz.). African Journal of Biotechnology, 8(15), p. 3424-3430. |
| spellingShingle | cassava nacl-ctab-azide solution phenol genogrinder Bhattacharjee, Ranjana Ferguson, Morag E. Gedil, Melaku A Dumet, D. Ingelbrecht, I.L. Field collection,preservation and large scale DNA extraction procedures for cassava (Manihot esculenta Crantz.) |
| title | Field collection,preservation and large scale DNA extraction procedures for cassava (Manihot esculenta Crantz.) |
| title_full | Field collection,preservation and large scale DNA extraction procedures for cassava (Manihot esculenta Crantz.) |
| title_fullStr | Field collection,preservation and large scale DNA extraction procedures for cassava (Manihot esculenta Crantz.) |
| title_full_unstemmed | Field collection,preservation and large scale DNA extraction procedures for cassava (Manihot esculenta Crantz.) |
| title_short | Field collection,preservation and large scale DNA extraction procedures for cassava (Manihot esculenta Crantz.) |
| title_sort | field collection preservation and large scale dna extraction procedures for cassava manihot esculenta crantz |
| topic | cassava nacl-ctab-azide solution phenol genogrinder |
| url | https://hdl.handle.net/10568/90807 |
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