title: Microsatellite (SSR) amplification by PCR usually led to polymorphic bands: Evidence which shows replication slippage occurs in extend or nascent DNA strands.
abstract: Microsatellites or simple sequence repeats (SSRs) are very effective molecular markers in population genetics, genome mapping, taxonomic study and other large-scale studies. Variation in number of tandem repeats within microsatellite refers to simple sequence length polymorphism (SSLP); but there are a few studies that are showed SSRs replication slippage may be occurred duringin vitroamplification which are produced 'stutter products' differing in length from the main products. The purpose of this study is introducing a reliable method to realize SSRs replication slippage. At first, three unique primers designed to amplify SSRs loci in the great gerbil (Rhombomys opimus) by PCR. Crush and soak method used to isolate interesting DNA bands from polyacrylamide gel. PCR products analyzed using by sequencing methods. Our study has been shown thatTaqDNA polymerase slipped during microsatellitein vitroamplification which led to insertion or deletion of repeats in sense or antisense DNA strands. It is produced amplified fragments with various lengths in gel electrophoresis showed as 'stutter bands'. Thus, in population studies by SSRs markers recommend that replication slippage effects and stutter bands have been considered.
authors.affiliation: Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Mazandaran, Iran.
authors.forename: Abasalt
authors.initials: A
authors.lastname: Hosseinzadeh-Colagar
authors.affiliation: Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Mazandaran, Iran.
authors.forename: Mohammad Javad
authors.initials: MJ
authors.lastname: Haghighatnia
authors.affiliation: Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Mazandaran, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Amiri
authors.affiliation: Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Mazandaran, Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Mohadjerani
authors.affiliation: Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Mazandaran, Iran.
authors.forename: Majid
authors.initials: M
authors.lastname: Tafrihi
keywords: Microsatellites; Replication slippage; Taq polymerase slippage