TY  - JOUR
AB  - Despite recent advances in high-throughput sequencing, difficulties are often encountered when developing microsatellites for species with large and complex genomes. This probably reflects the close association in many species of microsatellites with cryptic repetitive elements. We therefore developed a novel approach for isolating polymorphic microsatellites from the club-legged grasshopper (Gomphocerus sibiricus), an emerging quantitative genetic and behavioral model system. Whole genome shotgun Illumina MiSeq sequencing was used to generate over three million 300 bp paired-end reads, of which 67.75% were grouped into 40,548 clusters within RepeatExplorer. Annotations of the top 468 clusters, which represent 60.5% of the reads, revealed homology to satellite DNA and a variety of transposable elements. Evaluating 96 primer pairs in eight wild-caught individuals, we found that primers mined from singleton reads were six times more likely to amplify a single polymorphic microsatellite locus than primers mined from clusters. Our study provides experimental evidence in support of the notion that microsatellites associated with repetitive elements are less likely to successfully amplify. It also reveals how advances in high-throughput sequencing and graph-based repetitive DNA analysis can be leveraged to isolate polymorphic microsatellites from complex genomes.
DA  - 2016
DO  - 10.1002/ece3.2305
KW  - Acrididae
KW  - genetic marker development
KW  - Gomphocerus sibiricus
KW  - high-throughput sequencing
KW  - microsatellite
KW  - Orthoptera
KW  - transposable elements
LA  - eng
IS  - 6
M2  - 5718
PY  - 2016
SN  - 2045-7758
SP  - 5718-5727
T2  - Ecology and Evolution
TI  - High-throughput sequencing and graph-based cluster analysis facilitate microsatellite development from a highly complex genome
UR  - https://nbn-resolving.org/urn:nbn:de:0070-pub-29049896
Y2  - 2024-11-21T20:42:03
ER  -