Stamato T D, Hohmann L K
Cytogenet Cell Genet. 1975;15(6):372-9. doi: 10.1159/000130536.
A new technique of replica plating for CHO cells has been developed in which the pattern of colonies growing in a tissue culture plate is reproduced on a nylon cloth. The efficiency of replica plating onto the nylon cloth is affected by the colony size and the time of cloth-to-plate contact. Maximum replica plating efficiency (80-90%) occurs using 5- to 6-day-old colonies and a contact time of 4 h. The method easily allows 10(4) colonies to be examined for mutant phenotypes. The effectiveness of the technique is demonstrated by isolation of a clone deficient in activity of glucose-6-phosphate dehydrogenase.
已开发出一种用于中国仓鼠卵巢(CHO)细胞的复制平板接种新技术,该技术可将组织培养板中生长的菌落模式复制到尼龙布上。复制到尼龙布上的效率受菌落大小和布与板接触时间的影响。使用5至6日龄的菌落和4小时的接触时间可实现最大复制平板接种效率(80 - 90%)。该方法能够轻松检测10⁴个菌落的突变表型。通过分离缺乏葡萄糖-6-磷酸脱氢酶活性的克隆证明了该技术的有效性。