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After reading through your revised manuscript I see no reason to return it again to the referees. Both referees that made comments were supportive of publication following minor revisions.
I am satisfied that your revisions have addressed both my and the referees' comments. So I am happy to accept your manuscript and move it forward into press.
The referees agreed this short-note type manuscript, containing the details for microsatellite loci such as primer sequences, amplification conditions, polymorphism levels, should be accepted pending minor revisions. Overall, I believe that referees 1 and 2 have provided clear and specific advise on revision that will improve the manuscript. If you have any questions about it, please contact me.
In addition to the referees suggestions I would like you to consider the following:
line 79-81, maybe change sentence “Loci were in SLGD if known haploids produced a single allele and diploids produced either one or two alleles in their homozygous or heterozygous state, respectively” to line 77 just after “(Table 3, Supplementary Figure 1).”
line 87-89, the sentence “However, Krueger-Hadfield et al. (2013) demonstrated a strong bias in the maximum likelihood estimators of null allele frequency when macroalgal populations do
not undergo random mating.” probably fits better in the discussion.
line 94, “Wattier et al. (1998) demonstrated…” or developed?
line 98, which package which function in R, specifically?
I would also like to see some reference to other seaweed studies in which from a large pool of screened microsatellites, only a few revealed polymorphism. For instance, in three species of the genus Caulerpa for instance, three, seven and nine polymorphic microsatellites were uncovered [Varela-Álvarez et al, 2011, J.Appl. Phycol.]. The number of alleles in the present study is also low, raging from two to eleven. And to refer to the usefulness of this low level of polymorphism to successfully discriminate clonal lineages and analyse the clonal and genetic composition of algal beds. [Arnaud-Haond et al., 2013, Conservation genetics].
I look forward to seeing your revised manuscript so that I can move it forward into production.
I have no issues with this manuscript. Although the number of alleles detected across the polymorphic loci is rather low, the complete set should find some use in population genetic studies.
This technical note describes the development and characterization of microsatellite markers in an invasive estuarine rhodophyte exhibiting a cryptic biphasic life-cycle.
The theoretical and ecological relevance of the focal species are well framed and the potential usefulness of microsatellite markers to address a range of biogeographical, seascape and mating system related questions are pertinent.
I found the quality of writing good and the ideas expressed were very easy to follow. Manuscript structure appears to follow submission guidelines.
I did not find the necessary references to molecular data availability – the second column of Table 2 (Genbank accession numbers) must be filled, and the raw genotypic data for the 4 surveyed populations should also be made available in Dryad, Figshare or other repository.
I could not find any reference to possible funding sources.
Briefly, primers were developed for a selection of microsatellites previously identified in a contig library, and amplification, polymorphism, determinism and allele drop-out was assessed for each loci. A smaller set of polymorphic microsatellites were then used to characterize intra-specific patterns of diversity in a panel of geographically diverse populations.
I found the approach and analyses adequate for microsatellite characterization and selection. My only question is why the maximum number of microsatellite repeats would be set to 14 (page 4, line 55).
The use of diploid populations from each side of two different oceans and including native and alien populations predictably increased the amount of global variability detected.
Description of diversity within population was also adequate. Some additional analyses of these preliminary population data could allow readers to have an idea of the degree of differentiation between and within populations/ocean basins.
Page 4, Line 62: PCRs were performed using stock DNA or diluted DNA?
Page 4, Lines 63-65: nM is not clear, is this final concentration? maybe µM, mM?
Description of results was straightforward but sufficient, fully justifying the final selection of microsatellites.
I've uploaded an edited version of the original pdf with minor suggestions
The manuscript is nicely written and the methodological approach is appropriate. I find as limitation that the library was generated by a single individual. However having those primers tested on a bunch of individuals from 4 populations, may overcome (although not completely) the problem.
The authors report in the results (lines 117-120) that they have screened 42 loci, but only 9 were polymorphic, 13 monomorphic, 16 did not amplify and 4 had multipeaks. I suggest to compact table 2 keeping only the 9 polymorphic loci, move the13 monomorphic loci into a new supplementary table and discard all others as they have no use.
As final remark, the manuscript can be considered for publication after minor revisions.
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