REUSE OF DENATURING POLYACRYLAMIDE GELS FOR SHORT TANDEM REPEAT ANALYSIS

Citation
A. Tereba et al., REUSE OF DENATURING POLYACRYLAMIDE GELS FOR SHORT TANDEM REPEAT ANALYSIS, BioTechniques, 25(5), 1998, pp. 892-897
Citations number
18
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biology,"Biochemical Research Methods
Journal title
BioTechniques → ACNP
ISSN journal
0736-6205
Volume
25
Issue
5
Year of publication
1998
Pages
892 - 897
Database
ISI
SICI code
0736-6205(1998)25:5<892:RODPGF>2.0.ZU;2-6
Abstract
Denaturing polyacrylamide gel electrophoretic analysis of amplified po lymorphic short tandem repeat (STR) loci using fluorescent markers is a mainstay of forensic and paternity testing. To reduce the drawback o f preparing gels or using expensive precast gels, we have developed a simple and rapid method to reuse gels between 2 and 8 times over a per iod of several days. Following the initial electrophoresis and scan, t he original samples are removed from the gel by a 1-1.5-h reverse-elec trophoresis step. This step heats the gel for the next set of samples and can be performed several days after the initial electrophoresis. S ample bands remain sharp on subsequent runs, but edge effects (frownin g of the outside lanes) become progressively worse and ultimately limi t gel reuse. Well distortions and separation of the gel from the plate s become problems if the gel is used more than twice. However; degassi ng the gel solution and bonding the gel to both plates eliminate these problems. Precast gels also can be used multiple rimes. Using this te chnique, we have successfully analyzed samples amplified with a nine-l ocus multiplex system and characterized the separated products using a fluorescent scanner and software.