Background: Transgenic animals are widely used for research and for most of them, genotyping is unavoidable. Published protocols may be powerful but may also present disadvantages such as their cost or the requirement of additional steps/equipment. Moreover, if more than one strain must be genotyped, several protocols may need to be developed.
Methods: we adapted the existing amplification-resistant mutation protocol to develop the 1-h universal genotyping protocol (1-HUG), which allows the robust genotyping of genetically modified mice in 1 h from sample isolation to polymerase chain reaction gel running.
Results: This protocol allows the genotyping of different mouse models including mdx mouse, and FLExDUX4 and HSA-MerCreMer alone or in combination. It can be applied to different types of genomic modifications and to sexing.
Conclusions: The 1-HUG protocol can be used routinely in any laboratory using mouse models for neuromuscular diseases.
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http://dx.doi.org/10.1002/mus.26841 | DOI Listing |
J Inherit Metab Dis
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Department of Biomedical Science, The Graduate School, Kyungpook National University, Daegu, Republic of Korea.
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First Hospital of Quanzhou Affiliated to Fujian Medical University, Quanzhou, Fujian, China.
Despite the high sepsis-associated mortality, effective and specific treatments remain limited. Using conventional antibiotics as TIENAM (imipenem and cilastatin sodium for injection, TIE) is challenging due to increasing bacterial resistance, diminishing their efficacy and leading to adverse effects. We previously found that aloe-emodin (AE) exerts therapeutic effects on sepsis by reducing systemic inflammation and regulating the gut microbiota.
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