- Tail PCR Protocols (Preuss Lab, University of Chicago)
TAIL is a series of reactions that are intended to map where a T-DNA (transfer DNA) has inserted within the genome. The main components of the 3 reactions are the AD (Arbitrary Degenerate) primers, border primers, and DNA from the T-DNA lines that are to be mapped. AD primers are degenerate primers that anneal throughout the genome. The border primers are specific for the left and right borders of the T-DNA. From the primary reaction to the tertiary, the border primers get closer to the edge of the T-DNA. That is why a 'shift' is visible when running a gel with the secondary and tertiary reactions next to each other. The success rate of TAIL-PCR varies, depending onhow many DNA samples, AD primers, and border primers are used.
Added: Wed Feb 11 2009, Hits: 874, Reviews: 0
(Langdale Lab, Department of Plant Sciences, University of Oxford)
TAIL-PCR is a powerful tool for the recovery of DNA fragments adjacent to known sequences. Basically,
TAIL-PCR utilises three nested primers in consecutive reactions together with an arbitrary degenerate (AD)
primer having a lower Tm (melting temperature) so that the relative amplification frequencies of specific
and non-specific products can be thermally controlled.
Added: Wed Feb 11 2009, Hits: 2746, Reviews: 0
Tail PCR Protocols
(University of Florida)
Mu TAIL PCR produces a population of fragments with one end anchored in Robertson's Mutator (Mu) TIR's and the other in the Mu flanking DNA. These products may be used for cloning, gel analysis, or as hybridization probes. This protocol details the steps to take to improve the chances for uncontaminated and reproducible TAIL reactions.
Added: Wed Feb 11 2009, Hits: 809, Reviews: 0