KRISHI
ICAR RESEARCH DATA REPOSITORY FOR KNOWLEDGE MANAGEMENT
(An Institutional Publication and Data Inventory Repository)
"Not Available": Please do not remove the default option "Not Available" for the fields where metadata information is not available
"1001-01-01": Date not available or not applicable for filling metadata infromation
"1001-01-01": Date not available or not applicable for filling metadata infromation
Please use this identifier to cite or link to this item:
http://krishi.icar.gov.in/jspui/handle/123456789/6206| Title: | Polymerase chain reaction analysis of transgenic plants contaminated by Agrobacterium. |
| Other Titles: | Not Available |
| Authors: | Nain V, Jaiswal R, Dalal M, Ramesh B, Kumar PA. |
| ICAR Data Use Licennce: | http://krishi.icar.gov.in/PDF/ICAR_Data_Use_Licence.pdf |
| Author's Affiliated institute: | ICAR::National Research Centre on Plant Biotechnology |
| Published/ Complete Date: | 2005-03-01 |
| Project Code: | Not Available |
| Keywords: | Agrobacterium contamination, DNA renaturation, PCR, transgenic plants |
| Publisher: | Springer |
| Citation: | Not Available |
| Series/Report no.: | Not Available; |
| Abstract/Description: | Agrobacterium-mediated genetic transformation is a method of choice for the development of transgenic plants. The presence of latent Agrobacterium that multiplies in the plant tissue in spite of antibiotic application confounds the results obtained by polymerase chain reaction (PCR) analysis of putative transgenic plants. The presence of Agrobacterium can be confirmed by amplification of either Agrobacterium chromosomal genes or genes present out of transfer DNA (T-DNA) in the binary vector. However, the transgenic nature of Agrobacterium-contaminated transgenic plants cannot be confirmed by PCR. Here we report a simple protocol for PCR analysis of Agrobacterium-contaminated transgenic plants. This protocol is based on denaturation and renaturation of DNA. The contaminating plasmid vector becomes double-stranded after renaturation and is cut by a restriction enzyme having site(s) within the PCR amplicon. As a result, amplification by PCR is not possible. The genomic DNA with a few copies of the transgene remains single-stranded and unaffected by the restriction enzyme, leading to amplification by PCR. This protocol has been successfully tested with 4 different binary vectors and 3 Agrobacterium tumefaciens strains: EHA105, LBA4404, and GV3101. |
| Description: | Not Available |
| ISSN: | Not Available |
| Type(s) of content: | Research Paper |
| Sponsors: | Not Available |
| Language: | English |
| Name of Journal: | Plant Molecular Biology Reporter |
| NAAS Rating: | 7.34 |
| Volume No.: | 23 |
| Page Number: | 59-65 |
| Name of the Division/Regional Station: | Not Available |
| Source, DOI or any other URL: | https://link.springer.com/article/10.1007/BF02772647 |
| URI: | http://krishi.icar.gov.in/jspui/handle/123456789/6206 |
| Appears in Collections: | CS-NIPB-Publication |
Files in This Item:
There are no files associated with this item.
Items in KRISHI are protected by copyright, with all rights reserved, unless otherwise indicated.