Romain Joubert1, Virginie Mariot1, Julie Dumonceaux1. 1. NIHR Biomedical Research Centre, University College London, Great Ormond Street Institute of Child Health and Great Ormond Street Hospital NHS Trust, London, UK.
Abstract
INTRODUCTION: 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-hour universal genotyping protocol (1-HUG), which allows the robust genotyping of genetically modified mice in 1 h from sample isolation to PCR 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. DISCUSSION: The 1-HUG protocol can be used routinely in any laboratory using mouse models for neuromuscular diseases. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
INTRODUCTION: 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-hour universal genotyping protocol (1-HUG), which allows the robust genotyping of genetically modified mice in 1 h from sample isolation to PCR 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. DISCUSSION: The 1-HUG protocol can be used routinely in any laboratory using mouse models for neuromuscular diseases. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.