Isolation of High Molecular Weight DNA Suitable for BAC Library Construction from Woody Perennial Soft-Fruit Species
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| Title: | Isolation of High Molecular Weight DNA Suitable for BAC Library Construction from Woody Perennial Soft-Fruit Species |
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| Authors: | Hein, Ingo, Williamson, Sandie, Russell, Joanne, Powell, Wayne |
| Contributors: | School of Agriculture, Food and Wine |
| Source: | BioTechniques, Vol 38, Iss 1, Pp 69-71 (2005) |
| Publisher Information: | Informa UK Limited, 2005. |
| Publication Year: | 2005 |
| Subject Terms: | 0301 basic medicine, Chromosomes, Artificial, Bacterial, 0303 health sciences, DNA, Plant, QH301-705.5, Plant Leaves/genetics, Specimen Handling/methods, Bacterial/genetics, Ultrafiltration, Fractional Precipitation, DNA, Chromosomes, Ultrafiltration/methods, Specimen Handling, Molecular Weight, Plant Leaves, 03 medical and health sciences, Artificial, Plant/chemistry, Biology (General), Rosales, Rosales/genetics, Gene Library |
| Description: | We have developed a novel nuclei extraction method that allows for the extraction of high molecular weight DNA from leaves of woody perennial soft-fruit species that contain high levels of carbohydrates and polyphenolics. The method utilizes a modified buffer system including 4% (w/v) polyvinylpyrrolidone (PVP)-10 and a combination of nylon filters and Percoll gradients to purify nuclei extracts prior to embedding in agarose plugs. The effectiveness of the method was demonstrated on leaves of red raspberry (Rubus idaeus) and blackcurrant (Ribes nigrum), two soft-fruit species that have shown to be recalcitrant to standard genomic DNA extraction methods. Extracted DNA was readily digested by restriction enzymes and, as shown for raspberry, suitable for bacterial artificial chromosome (BAC) library construction. |
| Document Type: | Article |
| Language: | English |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/05381st02 |
| Access URL: | https://pubmed.ncbi.nlm.nih.gov/15679088 https://doaj.org/article/aaa9ddb0fccd482b8058b27279419a61 https://www.future-science.com/doi/pdf/10.2144/05381ST02 https://discovery.dundee.ac.uk/en/publications/isolation-of-high-molecular-weight-dna-suitable-for-bac-library-c https://www.future-science.com/doi/full/10.2144/05381ST02 http://www.biotechniques.com/article/05381ST02 https://www.ncbi.nlm.nih.gov/pubmed/15679088 https://dialnet.unirioja.es/servlet/articulo?codigo=3589103 |
| Rights: | CC BY CC BY NC ND |
| Accession Number: | edsair.doi.dedup.....3f28ceb44ee6e0defa8fe8c9856c2658 |
| Database: | OpenAIRE |
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Nájsť tento článok vo Web of Science | Abstract: | We have developed a novel nuclei extraction method that allows for the extraction of high molecular weight DNA from leaves of woody perennial soft-fruit species that contain high levels of carbohydrates and polyphenolics. The method utilizes a modified buffer system including 4% (w/v) polyvinylpyrrolidone (PVP)-10 and a combination of nylon filters and Percoll gradients to purify nuclei extracts prior to embedding in agarose plugs. The effectiveness of the method was demonstrated on leaves of red raspberry (Rubus idaeus) and blackcurrant (Ribes nigrum), two soft-fruit species that have shown to be recalcitrant to standard genomic DNA extraction methods. Extracted DNA was readily digested by restriction enzymes and, as shown for raspberry, suitable for bacterial artificial chromosome (BAC) library construction. |
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| ISSN: | 19409818 07366205 |
| DOI: | 10.2144/05381st02 |