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A Simple Method for Developing Microsatellite Markers using Amplified Fragments of Inter-simple Sequence Repeat (ISSR)
Authors:Chunlan Lian  Zhihua Zhou  Taizo Hogetsu
Institution:(1) Asian Natural Environmental Science Center, the University of Tokyo, 1–1–8 Midori-cho, Nishi-Tokyo, Tokyo, 188–0002 Japan, JP
Abstract:10 A primer IP1 designed from the sequenced region at one end of the microsatellite and for nested PCR another primer IP2 based on the sequence between IP1 and the microsatellite were prepared. These two primers were used to determine the other sequence flanking the microsatellite by a “walking” method. With this approach, we developed several microsatellite markers from Salix reinii, Pinus densiflora and Robinia pseudoacacia, respectively. The absence of enrichment processes and screening procedures makes it easier to develop microsatellite markers, and this approach provides an alternative for the development of microsatellite markers in any organism. Received 23 April 2001/ Accepted in revised form 11 June 2001
Keywords:: Inter-simple sequence repeat  Microsatellite isolation    Walking”  sequencing
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