R-ISSR as a new tool for genomic fingerprinting, mapping, and gene tagging |
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Authors: | Chunjiang Ye Zhanwang Yu Fanna Kong Suowei Wu Bin Wang |
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Institution: | (1) The State Key Laboratory for Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 100101 Beijing, P.R. China;(2) The Graduate School of Chinese Academy of Sciences, 100039 Beijing, P.R. China |
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Abstract: | In the present study we propose and test the concept of R-ISSR, a new tool for genomic fingerprinting, mapping, and gene tagging.
The concept is based on the fact that primers for inter-simple sequence repeat (ISSR) and random-amplified polymorphic DNA
(RAPD) analysis elicit different genomic information, and the combined use of these 2 kinds of primers in the same polymerase
chain reaction (PCR) reactions might reveal new genomic loci that could not be detected with either technique alone. The feasibility
of this tool was first electronically simulated with sequence analysis software andArabidopsis chromosome sequence. Next, different combinations of ISSR and RAPD primers were applied in real PCR reactions to detect new
genomic loci in 2 maize lines (Q319 and 1145). Sequencing gels were used to separate PCR products and showed good resolving
ability in comparison with agarose gels. RAPD primers could be successfully used with ISSR primers for the detection of new
genomic loci and applied in a new way for genomic mapping, fingerprinting, and gene tagging. |
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Keywords: | gene tagging ISSR RAPD sequencing gel SSR |
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