Cloning flanking sequence by single-primer PCR in transgenic plants
نویسندگان
چکیده
منابع مشابه
Cloning flanking sequence by single-primer PCR in transgenic plants.
The insertion position of exogenous genes in plant genomes is usually identified by adapter ligation-mediated polymerase chain reaction (PCR), thermal asymmetric interlaced PCR, and restriction site extension PCR in transgenic plant research. However, these methods have various limitations, such as the complexity of designing primers and time-consuming and multiple-step procedures. The goal of ...
متن کاملCloning of flanking sequence in transgenic plants by restriction site-anchored single-primer polymerase chain reaction.
Determining the insertion position of an exogenous gene in the target plant genome is one of the main issues in the transgenic plant field. This study introduced a simple, rapid, and accurate method to clone the flanking sequences of the transgenic bar gene as the anchoring gene in the transgenic maize genome using single-primer polymerase chain reaction (PCR). This method was based on the dist...
متن کاملSingle-primer amplification of flanking sequences.
“megaprimer method” of site-directed mutagenesis. BioTechniques 8:404-407. 11.Senanayake, S.D. and D.A. Brian. 1995. Precise large deletions by the PCR based overlap extention. Mol. Biotechnol. 4:13-15. 12.Silver, J., T. Limjoco, and Feinstone. 1995. Site-specific mutagenesis using the polymerase chain reaction, p. 179-188. In M.A. Innis, D.H. Gelfand and J.J. Sninsky (Eds.), PCR Strategies. Ac...
متن کاملBiologically-generated primer for PCR: PCR primer of unknown sequence
We describe a method for producing specific PCR primers directly from PCR product, bypassing the usual need to know the primer sequence. Lack of abundance of primers derived from a PCR product is compensated for by the incorporation of an arbitrary 5'TAG sequence which acts as a surrogate template target for the bulk amplification phase. We use the technique to amplify clonospecific rearranged ...
متن کاملFusion primer and nested integrated PCR (FPNI-PCR): a new high-efficiency strategy for rapid chromosome walking or flanking sequence cloning
BACKGROUND The advent of genomics-based technologies has revolutionized many fields of biological enquiry. However, chromosome walking or flanking sequence cloning is still a necessary and important procedure to determining gene structure. Such methods are used to identify T-DNA insertion sites and so are especially relevant for organisms where large T-DNA insertion libraries have been created,...
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ژورنال
عنوان ژورنال: Genetics and Molecular Research
سال: 2014
ISSN: 1676-5680
DOI: 10.4238/2014.october.20.16