Amplification of genomic sequences flanking T-DNA insertions by thermal asymmetric interlaced polymerase chain reaction

Methods Mol Biol. 2005;286:341-8. doi: 10.1385/1-59259-827-7:341.

Abstract

Thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR) is an efficient tool for the recovery of DNA fragments adjacent to known sequences. A protocol is presented for the amplification of genomic sequences flanking DNA (T-DNA or transposon) insertions using the TAIL-PCR method. The amplified products are suitable as templates for direct sequencing, or for cloning in vectors. Examples are given in use of the protocol for T-DNA tagging in rice using the pCAMBIA binary vectors.

MeSH terms

  • Base Sequence
  • DNA Primers
  • DNA, Bacterial / genetics*
  • Genes, Plant / genetics*
  • Genetic Vectors
  • Molecular Sequence Data
  • Plants, Genetically Modified / genetics*
  • Polymerase Chain Reaction / methods*
  • Thermodynamics

Substances

  • DNA Primers
  • DNA, Bacterial
  • T-DNA