NDLI: Frequent problems and their resolutions by using thermal asymmetric interlaced PCR (TAIL-PCR) to clone genes in Arabidopsis T-DNA tagged mutants

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Frequent problems and their resolutions by using thermal asymmetric interlaced PCR (TAIL-PCR) to clone genes in Arabidopsis T-DNA tagged mutants

Content Provider	Semantic Scholar
Author	Zhang, Dan Song, Bin Guo, Guang-Qin
Copyright Year	2015
Abstract	T-DNA insertional mutagenesis is a powerful tool in Arabidopsis functional genomics research. Previous studies have developed thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR) as an efficient strategy in isolation of DNA sequences adjacent to known sequences in T-DNA tagged mutants. However, a number of problems are encountered when attempts are made to clone flanking sequences in T-DNA tagged mutants. Therefore, it is necessary to improve the efficiency of cloning mutagenesis. Here, we present the most frequent problems and provide an improved method to increase TAIL-PCR efficiency. Even then, it is not always possible to successfully obtain flanking sequences; in such cases, we recommend using high-throughput sequencing to determine the mutations.
Starting Page	260
Ending Page	267
Page Count	8
File Format	PDF HTM / HTML
PubMed reference number	26019639v1
Volume Number	29
Journal	Biotechnology, biotechnological equipment
Language	English
Access Restriction	Open
Subject Keyword	Biological Science Disciplines Biopolymer Sequencing Checking (action) Clone Clone Cells Flank (surface region) Gene Knockout Techniques Genomics Insertion Mutation Instrument - device Language Development Disorders Mandibular right second molar tooth Mutagenesis, Insertional Mutation Nucleic Acid Hybridization Phenotype Polymerase Chain Reaction Reverse Transcriptase Polymerase Chain Reaction Tail Tracer Wu-ling-San hachimijiogan mutant
Content Type	Text
Resource Type	Article

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