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目的:利用错配PCR鉴别小片段缺失的纯合子与野生型斑马鱼。方法:将突变型斑马鱼杂合子F1杂交,收集胚胎裂解获得粗基因组模板,进行第1次PCR基因分型,经凝胶电泳后确认,对单条带产物的模板进行第2次PCR基因分型(利用错配PCR引物),再次经凝胶电泳检测。结果:利用错配PCR引物进行基因分型,纯合子PCR产物为单一条带,而野生型PCR产物无条带。结论:该方法稳定可靠,省时省钱,可用于小片段缺失的纯合子与野生型斑马鱼的鉴别。
OBJECTIVE: To identify small fragments of homozygous and wild-type zebrafish using mismatched PCR. Methods: The mutant zebrafish hybrid F1 was hybridized and the embryos were lysed to obtain the crude genome template. The first PCR genotyping was performed and confirmed by gel electrophoresis. The second PCR gene was performed on the template of the single band product (Using mismatched PCR primers) and again detected by gel electrophoresis. RESULTS: Genotyping was performed using mismatched PCR primers, homozygous PCR products were single band, whereas wild type PCR products were unlabeled. Conclusion: The method is stable and reliable, saves time and money, and can be used for the identification of small homozygotes and wild-type zebrafish.