A Simple Method for Developing Microsatellite Markers using Amplified Fragments of Inter-simple Sequence Repeat (ISSR)

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.