Schmitteckert E M, Prokop C M, Hedrich H J
Institute of Pharmacology and Toxicology, University of Wuerzburg, Germany.
Lab Anim. 1999 Oct;33(4):385-9. doi: 10.1258/002367799780487922.
The breeding of transgenic animals requires that each individual offspring be analysed for integration of transgenic deoxyribonucleic acid (DNA), unless exclusively homozygous animals are mated. The standard protocol for identification of transgenic animals (Hogan et al. 1994) is based on tissue samples and preparation of chromosomal DNA including proteinase K digestion and phenol/chloroform extraction. The procedure described here represents a much simpler and faster method to screen offspring for the transgene DNA. It is based on the use of hair bulbs as sample material, which can be directly used for polymerase chain reaction (PCR) after alkaline lysis. This protocol allows large numbers of animals to be easily screened in a minimum amount of time. A unique advantage though, is the reduction of the distress caused to the animals. With respect to the 3Rs (Replacement, Reduction, Refinement), and because of technical advantages this method may replace ear or tail clipping.
转基因动物的培育要求对每一个后代进行转基因脱氧核糖核酸(DNA)整合分析,除非只交配纯合动物。鉴定转基因动物的标准方案(霍根等人,1994年)基于组织样本和染色体DNA的制备,包括蛋白酶K消化和酚/氯仿提取。这里描述的程序是一种筛选后代转基因DNA的简单得多且更快的方法。它基于使用毛球作为样本材料,经碱性裂解后可直接用于聚合酶链反应(PCR)。该方案能在最短时间内轻松筛选大量动物。不过,一个独特的优势是减少了对动物造成的痛苦。就3R原则(替代、减少、优化)而言,由于技术优势,这种方法可能会取代剪耳或剪尾。