Next Article in Journal
Previous Article in Journal
Int. J. Mol. Sci. 2012, 13(6), 7586-7593; doi:10.3390/ijms13067586
Article

Isolation and Characterization of Simple Sequence Repeats (SSR) Markers from the Moss Genus Orthotrichum Using a Small Throughput Pyrosequencing Machine

1,* , 2
, 1
, 2
, 1
 and 3
Received: 10 May 2012; in revised form: 12 June 2012 / Accepted: 13 June 2012 / Published: 19 June 2012
View Full-Text   |   Download PDF [138 KB, uploaded 19 June 2014]
Abstract: Here, we report the results of next-generation sequencing on the GS Junior system to identify a large number of microsatellites from the epiphytic moss Orthotrichum speciosum. Using a combination of a total (non-enrichment) genomic library and small-scale 454 pyrosequencing, we determined 5382 contigs whose length ranged from 103 to 5445 bp. In this dataset we identified 92 SSR (simple sequence repeats) motifs in 89 contigs. Forty-six of these had flanking regions suitable for primer design. We tested PCR amplification, reproducibility, and the level of polymorphism of 46 primer pairs for Orthotrichum speciosum using 40 individuals from two populations. As a result, the designed primers revealed 35 polymorphic loci with more than two alleles detected. This method is cost- and time-effective in comparison with traditional approaches involving cloning and sequencing.
Keywords: SSR markers; Orthotrichum; 454 pyrosequencing SSR markers; Orthotrichum; 454 pyrosequencing
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Export to BibTeX |
EndNote


MDPI and ACS Style

Sawicki, J.; Kwaśniewski, M.; Szczecińska, M.; Chwiałkowska, K.; Milewicz, M.; Plášek, V. Isolation and Characterization of Simple Sequence Repeats (SSR) Markers from the Moss Genus Orthotrichum Using a Small Throughput Pyrosequencing Machine. Int. J. Mol. Sci. 2012, 13, 7586-7593.

AMA Style

Sawicki J, Kwaśniewski M, Szczecińska M, Chwiałkowska K, Milewicz M, Plášek V. Isolation and Characterization of Simple Sequence Repeats (SSR) Markers from the Moss Genus Orthotrichum Using a Small Throughput Pyrosequencing Machine. International Journal of Molecular Sciences. 2012; 13(6):7586-7593.

Chicago/Turabian Style

Sawicki, Jakub; Kwaśniewski, Mirosław; Szczecińska, Monika; Chwiałkowska, Karolina; Milewicz, Monika; Plášek, Vítězslav. 2012. "Isolation and Characterization of Simple Sequence Repeats (SSR) Markers from the Moss Genus Orthotrichum Using a Small Throughput Pyrosequencing Machine." Int. J. Mol. Sci. 13, no. 6: 7586-7593.


Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert