Microfluidic-aided genotyping of zebrafish in the first 48 h with 100% viability.
Microfluidic-aided genotyping of zebrafish in the first 48 h with 100% viability.
复制标题
DOI：
10.1007/s10544-015-9946-9
复制
发表时间：
2015-04
影响因子：
2.8
通讯作者：
Gale BK
中科院分区：
文献类型：
--
作者：
Samuel R;Stephenson R;Roy P;Pryor R;Zhou L;Bonkowsky JL;Gale BK
本文介绍了一种新的斑马鱼胚胎基因分型方法，在不牺牲胚胎生命/健康的情况下，对1-2天大的斑马鱼胚胎进行基因分型。该方法利用微流控技术从胚胎的绒毛膜液或鳍组织中提取和收集少量遗传物质。然后，使用常规DNA提取、PCR扩增、高分辨率熔融分析和荧光DNA检测技术，对胚胎进行基因分型。绒毛膜液法成功率为78%，剪鳍法成功率为100%。绒毛膜液被证明只含有来自胚胎的DNA，而不含有来自母亲的DNA。这些结果提示了一种新的斑马鱼胚胎分型方法，这种方法可以促进高通量筛选，同时保持胚胎100%的存活率。
This paper introduces an innovative method for genotyping 1–2 days old zebrafish embryos, without sacrificing the life/health of the embryos. The method utilizes microfluidic technology to extract and collect a small amount of genetic material from the chorionic fluid or fin tissue of the embryo. Then, using conventional DNA extraction, PCR amplification, and high resolution melt analysis with fluorescent DNA detection techniques, the embryo is genotyped. The chorionic fluid approach was successful 78 % of the time while the fin clipping method was successful 100 % of the time. Chorionic fluid was shown to only contain DNA from the embryo and not from the mother. These results suggest a novel method to genotype zebrafish embryos that can facilitate high-throughput screening, while maintaining 100 % viability of the embryo.