Microsatellite marker identification using genome screening and restriction-ligation

We have identified a fast and easy method for finding microsatellite markers that utilizes genome screening with inter-simple sequence repeat (ISSR) primers to detect microsatellite regions and to obtain sequence information flanking one side of the microsatellites and a restriction-ligation techniq...

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Main Authors: Helena Korpelainen, Kirsi Kostamo, Viivi Virtanen
Format: Article
Language:English
Published: Taylor & Francis Group 2007-04-01
Series:BioTechniques
Online Access:https://www.future-science.com/doi/10.2144/000112415
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author Helena Korpelainen
Kirsi Kostamo
Viivi Virtanen
author_facet Helena Korpelainen
Kirsi Kostamo
Viivi Virtanen
author_sort Helena Korpelainen
collection DOAJ
description We have identified a fast and easy method for finding microsatellite markers that utilizes genome screening with inter-simple sequence repeat (ISSR) primers to detect microsatellite regions and to obtain sequence information flanking one side of the microsatellites and a restriction-ligation technique with a specific adaptor to allow sequence walking to obtain sequence information flanking the other side of the microsatellites. Two main alternatives of the method (with or without cloning) are presented. We successfully utilized the method when identifying microsatellite markers for 21 bryophyte species, three algal species, and for the raccoon dog. The proportion of polymorphic markers equaled 95%. We observed that microsatellites are commonly found within the sequenced ISSR amplification products (54% in the present study), in which case specific primers can be identified for the microsatellite without a further restriction-ligation step. It is evident that the DNA regions amplified by ISSR markers commonly represent microsatellite hotspots. We propose that the identified method and the knowledge of the common presence of additional microsatellite repeats within ISSR amplification products are especially attractive to researchers who conduct small-scale microsatellite identification, such as researchers in population genetics and conservation biology.
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spelling doaj-art-a3bf690ea10e465482a486ed488d27e52025-08-20T02:26:06ZengTaylor & Francis GroupBioTechniques0736-62051940-98182007-04-0142447948610.2144/000112415Microsatellite marker identification using genome screening and restriction-ligationHelena Korpelainen0Kirsi Kostamo1Viivi Virtanen21University of Helsinki, Helsinki, Finland1University of Helsinki, Helsinki, Finland1University of Helsinki, Helsinki, FinlandWe have identified a fast and easy method for finding microsatellite markers that utilizes genome screening with inter-simple sequence repeat (ISSR) primers to detect microsatellite regions and to obtain sequence information flanking one side of the microsatellites and a restriction-ligation technique with a specific adaptor to allow sequence walking to obtain sequence information flanking the other side of the microsatellites. Two main alternatives of the method (with or without cloning) are presented. We successfully utilized the method when identifying microsatellite markers for 21 bryophyte species, three algal species, and for the raccoon dog. The proportion of polymorphic markers equaled 95%. We observed that microsatellites are commonly found within the sequenced ISSR amplification products (54% in the present study), in which case specific primers can be identified for the microsatellite without a further restriction-ligation step. It is evident that the DNA regions amplified by ISSR markers commonly represent microsatellite hotspots. We propose that the identified method and the knowledge of the common presence of additional microsatellite repeats within ISSR amplification products are especially attractive to researchers who conduct small-scale microsatellite identification, such as researchers in population genetics and conservation biology.https://www.future-science.com/doi/10.2144/000112415
spellingShingle Helena Korpelainen
Kirsi Kostamo
Viivi Virtanen
Microsatellite marker identification using genome screening and restriction-ligation
BioTechniques
title Microsatellite marker identification using genome screening and restriction-ligation
title_full Microsatellite marker identification using genome screening and restriction-ligation
title_fullStr Microsatellite marker identification using genome screening and restriction-ligation
title_full_unstemmed Microsatellite marker identification using genome screening and restriction-ligation
title_short Microsatellite marker identification using genome screening and restriction-ligation
title_sort microsatellite marker identification using genome screening and restriction ligation
url https://www.future-science.com/doi/10.2144/000112415
work_keys_str_mv AT helenakorpelainen microsatellitemarkeridentificationusinggenomescreeningandrestrictionligation
AT kirsikostamo microsatellitemarkeridentificationusinggenomescreeningandrestrictionligation
AT viivivirtanen microsatellitemarkeridentificationusinggenomescreeningandrestrictionligation