一小时通用小鼠基因型鉴定方案。
One-hour universal protocol for mouse genotyping.
机构信息
NIHR Biomedical Research Centre, University College London, Great Ormond Street Institute of Child Health and Great Ormond Street Hospital NHS Trust, London, United Kingdom.
出版信息
Muscle Nerve. 2020 Jun;61(6):801-807. doi: 10.1002/mus.26841. Epub 2020 Mar 6.
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.
背景
转基因动物被广泛应用于研究，对于大多数动物来说，基因分型是不可避免的。已发表的方案可能很强大，但也可能存在一些缺点，如成本较高或需要额外的步骤/设备。此外，如果需要对多个品系进行基因分型，则可能需要开发多个方案。
方法
我们对现有的抗扩增突变方案进行了改编，开发了 1 小时通用基因分型方案（1-HUG），该方案允许从样本分离到聚合酶链反应凝胶运行，在 1 小时内对基因修饰的小鼠进行稳健的基因分型。
结果
该方案可对不同的小鼠模型进行基因分型，包括 mdx 小鼠、FLExDUX4 和 HSA-MerCreMer 单独或组合使用。它可应用于不同类型的基因组修饰和性别鉴定。
结论
1-HUG 方案可在任何使用神经肌肉疾病小鼠模型的实验室中常规使用。
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