Abstract:
:Terminal Restriction Fragment Length Polymorphism (T-RFLP) or Fluorescent Polymerase Chain Reaction/Restriction Fragment Length Polymorphism (FluRFLP) have made a significant impact on the way in which PCR products amplified from mixed community DNA extracts have been assessed. Technically, these approaches are essentially the same. PCR products are generated that contain at one 5' end label, typically a fluorescent moiety, that will be detected by a DNA sequencing machine. Upon digestion using a specific restriction endonuclease, labeled and unlabeled fragments are generated. This restriction endonuclease is chosen such that following this digestion, each labeled fragment corresponds to a different sequence variant. During electrophoretic separation, the DNA sequencing machine detects only these labeled fragments and therefore detects only the sequence variants. The aim of this article is to describe the protocols and demonstrate that this profiling can be performed using different DNA sequencing machines. The analysis and applications of this approach are also discussed.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Bruce KD,Hughes MRdoi
10.1385/MB:16:3:261keywords:
subject
Has Abstractpub_date
2000-11-01 00:00:00pages
261-9issue
3eissn
1073-6085issn
1559-0305pii
MB:16:3:261journal_volume
16pub_type
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