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    Field collection,preservation and large scale DNA extraction procedures for cassava (Manihot esculenta Crantz.)

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    U09ArtBhattachrjeeFieldNothomNodev.pdf (755.0Kb)
    Date
    2009
    Author
    Bhattacharjee, R.
    Ferguson, M.
    Gedil, M.
    Dumet, D.
    Ingelbrecht, I.L.
    Type
    Journal Article
    Metadata
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    Abstract/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.
    Permanent link to this item
    https://hdl.handle.net/20.500.12478/2839
    IITA Subjects
    Cassava; Genetic Improvement; Tissue Culture; Markets
    Agrovoc Terms
    Cassava; Nacl-Ctab-Azide Solution; Phenol; Genogrinder 2000
    Regions
    Africa; West Africa; East Africa
    Countries
    Nigeria; Kenya
    Collections
    • Journal and Journal Articles4835
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