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However, the general genotyping methods are time-consuming and unreliable given the loxP site is only 34\u00a0bp long. Here, based on the tetra primer-paired PCR amplification, we describe an efficient genotyping method which can simultaneously generate the internal control band, wild type (wt)-genotype band, and\/or loxP-genotype band through one single PCR amplification. It is easy to interpret the mouse genotypes from the pattern of the bands. Further, the results could also help to exclude the possibility of minor cross-contamination, since the ratio between the bands\u2019 quantity in wt\/wt, wt\/loxP, and loxP\/loxP mice are relatively constant, which makes the genotyping more reliable when it is performed in a large amount.<\/jats:p>","DOI":"10.1007\/s12033-022-00500-5","type":"journal-article","created":{"date-parts":[[2022,5,3]],"date-time":"2022-05-03T05:02:40Z","timestamp":1651554160000},"page":"1227-1233","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["One-Step Genotyping Method in loxP-Based Conditional Knockout Mice Generated by CRISPR-Cas9 Technology"],"prefix":"10.1007","volume":"64","author":[{"given":"He","family":"Zhu","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Siqian","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wenxi","family":"He","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fei","family":"Sun","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yang","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ping","family":"Yang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qilin","family":"Yu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8521-7916","authenticated-orcid":false,"given":"Shu","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2022,5,3]]},"reference":[{"key":"500_CR1","doi-asserted-by":"publisher","first-page":"5166","DOI":"10.1073\/pnas.85.14.5166","volume":"85","author":"B Sauer","year":"1988","unstructured":"Sauer, B., & Henderson, N. (1988). Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1. Proceedings of the National Academy of Sciences of the United States of America, 85, 5166\u20135170.","journal-title":"Proceedings of the National Academy of Sciences of the United States of America"},{"key":"500_CR2","doi-asserted-by":"publisher","first-page":"347","DOI":"10.1038\/nbt.2842","volume":"32","author":"JD Sander","year":"2014","unstructured":"Sander, J. D., & Joung, J. K. (2014). CRISPR-Cas systems for editing, regulating and targeting genomes. Nature Biotechnology, 32, 347\u2013355.","journal-title":"Nature Biotechnology"},{"key":"500_CR3","doi-asserted-by":"publisher","first-page":"e2000650","DOI":"10.1002\/biot.202000650","volume":"16","author":"Y Zhao","year":"2021","unstructured":"Zhao, Y., Karan, R., & Altpeter, F. (2021). Error-free recombination in sugarcane mediated by only 30 nucleotides of homology and CRISPR\/Cas9 induced DNA breaks or Cre-recombinase. 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