Abstract
In this protocol, determination of seed paternity by microsatellite markers in Nicotiana attenuata is described. However, this does not include a protocol for the novel marker selection/identification, but rather exploits the markers generated for a closely related species N. tabacum (Bindler et al., 2007). This is a high-throughput protocol optimized and streamlined for one skilled person to process 384 (96 x 4) seeds in 5 days, from DNA isolation (from seedlings) to paternity assessment by microsatellite genotype data.
Keywords: Microsatellite genotyping, Multiplex PCR, High-throughput seed paternity test, Nicotiana attenuata
Materials and Reagents
Equipment
Procedure
Recipes
Acknowledgments
This work was supported by the Max Planck Gesellschaft. The protocol was adapted from the publication: Kessler et al. (2012).
References
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.