ARS | Publication request: INTER-SIMPLE SEQUENCE REPEAT RESTRICTION FRAGMENT LENGTH POLYMORPHISMS FOR DNA FINGERPRINTING

Submitted to: Biotechniques
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: March 6, 2003
Publication Date: May 7, 2003
Citation: SYLVESTER, H.A. INTER-SIMPLE SEQUENCE REPEAT RESTRICTION FRAGMENT LENGTH POLYMORPHISMS FOR DNA FINGERPRINTING. BIOTECHNIQUES. 2003. Vol 34, No. 5, pgs. 942-944.

Interpretive Summary: This paper describes a new method of detecting DNA variation, using honey bees as an example. It presents a method of detecting DNA sequence variation that is sampler than using a DNA sequencer and that can detect DNA sequence variation in fragments of DNA that are too large to be detected in a DNA sequencer. This provides a new way to find DNA markers for breeding and selection experiments, particularly for identifying Primorsky honey bees imported by USDA, ARS from far-eastern Russia.

Technical Abstract: A simple method of DNA fingerprinting is described, where increased levels of genetic variation can be detected in PCR fragments too large to detect in a DNA sequencer, as well as in PCR fragments detectable in a DNA sequencer. Single microsatellite-anchored primers are used for PCR amplification of inter-simple sequence repeat fragments (ISSRs). The PCR products are digested with restriction enzymes and the restriction fragments compared in an electrophoresis gel. High levels of intraspecific polymorphism among restriction fragments were detected.