Multiplex Detection of SNPs for Genetic Monitoring in Laboratory Mice by Luminex xTAG Assay
, Jie Wei 1,†, Hong Wang 1, Huan Li 1, Lan Zhao 1, Rui Fu 1,* and Bingfei Yue 3,*Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe article show the importance of genetic quality in laboratory mice and their importance in biomedical reserach. So, this is important maintance the quality control of the background of the mice. In this study the research usie the Luminex XTAG method to improve the genetic monitoring practies and ensure better research quality.
This article is clear, basic in some parts with the deficency in introduction to regarding the background of the mice and their breeding. There are some grammatical errors.
The article need some simple modification:
- line 37: missing the point (.)
- line 39: remove point (.)
- line 41: remove double point (.)
- introduction: Add more details on the background of mice. The difference between inbred or outbred mice.
- line 112: W writing in tiny
- line 167: remove the (,) and add (.)
- line 173: the photo caption is too close to the text
- line 181: like line 173
- line 200: remove (.)
Comments for author File: 
Comments.pdf
Author Response
Comments 1:-line 37: missing the point (.)
Response 1: Thank you for pointing this out. We have add the point in it.
Comments 2:- line 39: remove point (.)
Response 2: Thank you for pointing this out. We have removed the point.
Comments 3:- line 41: remove double point (.)
Response 3: Thank you for pointing this out. We have removed one point.
Comments 4:- introduction: Add more details on the background of mice. The difference between inbred or outbred mice.
Response 4: Thank you for pointing this out. We agree with this comment. Therefore, We have added the difference between inbred mice and coutbred mice in lines 29 to 32.
Comments 5: - line 112: W writing in tiny
Response 5: Thank you for pointing this out. We have written W in tiny.
Comments 6: - line 167: remove the (,) and add (.)
Response 6: Thank you for pointing this out. We have removed the (,) and add (.).
Comments 7: - line 173: the photo caption is too close to the text
Response 7: Thank you for pointing this out. We have adjusted the line spacing.
Comments 8: - line 181: like line 173
Response 8: Thank you for pointing this out. We have adjusted the line spacing like comments 7.
Comments 9: - line 200: remove (.)
Response 9: Thank you for pointing this out. We have removed the (.) .
Reviewer 2 Report
Comments and Suggestions for AuthorsGenetic quality assurance, including genetic monitoring of mouse strains and background characterization of genetically altered mouse models, should be an important component of a genetic quality control program for genetic mouse model studies. In this study, the authors attempted to develop an efficient, high-throughput, low-cost, and universally applicable Luminex xTAG assay that would provide a good option for genetic monitoring of inbred and outbred mice. Overall, this is a good topic and research point, but there are still many issues that need to be clarified and confirmed by the authors.
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1. How do novel SNP marker panels based on allele-specific primer extension (ASPE) and MagPlex-TAG microsphere methods improve assay accuracy, time, and price? The authors need to provide more comparative details on how the novel method compares to existing and older methods. For example, create a summary table comparing each method from different points.
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2. What's the PCR cycle number in method 2.4?
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3. The authors state in the manuscript that they obtained 34 loci that could be stably amplified in inbred lines and 15 loci with good polymorphism in outbred groups. References are needed to support and confirm these loci.
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4. In Table 5, it is advisable to highlight the comparison region/bp in a different color so that it is easy for the reader to read.
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5. The authors note that “inbred mice from Beijing, Shanghai and Shenzhen were selected for this study.” So where are the Shenzhen mice? Why were the 34 loci randomized into 4 groups?
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6. “Examples of such corrected data for the 2 inbred strains (C57BL/6JNifdc and BALB/cJNifdc) used to develop this assay are shown in the Table 4”. Clarification is needed as to why only these two strains were chosen.
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7. Table 4 is very confusing, some of the SNP locus datasets are double datasets, e.g. rs3709624, and there are two different rows for each SNP locus dataset. The authors need to provide more information for this.
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8. Please describe the detailed formulas (for example, Ne, SI, PIC, Ave Het...) for calculating the data in Table 6. like, The Shannon-index (SI) has been used to estimate genetic diversity in numerous studies. The Shannon diversity index is calculated by taking the sum of Pi * ln(Pi) for each species. If more numbers are added based on the number of species present (or richness), then higher richness should increase the Shannon diversity index if the relative species abundance remains the same. (PI: The proportion of individuals of each species).
Authors are advised to review the manuscript again carefully to check for typos, grammatical errors, and redundant sentences for ease of readng.
Author Response
Please see the attachment.Author Response File: Author Response.pdf
Round 2
Reviewer 2 Report
Comments and Suggestions for AuthorsThe manuscript is a significant improvement over the previous manuscript in terms of writing and data arrangement. The authors responded appropriately to most of the issues raised by the reviewers. However, the formatting of the figures and tables in this review manuscript needs to be reorganized to comply with the requirements of the journal.
Author Response
Thank you for pointing this out. We have reorganized the tables in the article according to the requirements of the journal.
