Sample preparation for 2b-RAD genotyping
High-throughput sequencing of restriction site-associated genomic DNA (RAD) is now a widely used approach for simultaneously discovering and profiling genetic polymorphisms. We describe an alternative approach for RAD genotyping with several useful features, called 2b-RAD for the use of type IIB restriction endonucleases (AlfI, BsaXI). The fragments excised by type IIB enzymes are uniform in length, streamlining library preparation and producing even sequencing coverage across the genome. The cohesive ends present on both 5’ and 3’ ends of each fragment make it possible to target a defined subset of restriction sites using modified adaptors. Marker density can be adjusted as needed in this way, greatly reducing sequencing coverage requirements and costs. 2b-RAD offers a cost-effective strategy for high-throughput genotyping that is widely suitable for linkage mapping and profiling genetic variation in natural populations.
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Posted 06 Jun, 2012
Sample preparation for 2b-RAD genotyping
Posted 06 Jun, 2012
High-throughput sequencing of restriction site-associated genomic DNA (RAD) is now a widely used approach for simultaneously discovering and profiling genetic polymorphisms. We describe an alternative approach for RAD genotyping with several useful features, called 2b-RAD for the use of type IIB restriction endonucleases (AlfI, BsaXI). The fragments excised by type IIB enzymes are uniform in length, streamlining library preparation and producing even sequencing coverage across the genome. The cohesive ends present on both 5’ and 3’ ends of each fragment make it possible to target a defined subset of restriction sites using modified adaptors. Marker density can be adjusted as needed in this way, greatly reducing sequencing coverage requirements and costs. 2b-RAD offers a cost-effective strategy for high-throughput genotyping that is widely suitable for linkage mapping and profiling genetic variation in natural populations.
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