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In Ovo Electroporations of HH Stage 10 Chicken Embryos

机译:HH 10期鸡胚的卵内电穿孔

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摘要

Large size and external development of the chicken embryo have long made it a valuable tool in the study of developmental biology. With the advent of molecular biological techniques, the chick has become a useful system in which to study gene regulation and function. By electroporating DNA or RNA constructs into the developing chicken embryo, genes can be expressed or knocked down in order to analyze in vivo gene function. Similarly, reporter constructs can be used for fate mapping or to examine putative gene regulatory elements. Compared to similar experiments in mouse, chick electroporation has the advantages of being quick, easy and inexpensive. This video demonstrates first how to make a window in the eggshell to manipulate the embryo. Next, the embryo is visualized with a dilute solution of India ink injected below the embryo. A glass needle and pipette are used to inject DNA and Fast Green dye into the developing neural tube, then platinum electrodes are placed parallel to the embryo and short electrical pulses are administered with a pulse generator. Finally, the egg is sealed with tape and placed back into an incubator for further development. Additionally, the video shows proper egg storage and handling and discusses possible causes of embryo loss following electroporation.
机译:长期以来,鸡胚的大尺寸和外部发育一直使其成为研究发育生物学的宝贵工具。随着分子生物学技术的出现,雏鸡已成为研究基因调控和功能的有用系统。通过将DNA或RNA构建体电穿孔到发育中的鸡胚中,可以表达或敲除基因,以便分析体内基因功能。类似地,报告基因构建体可用于命运作图或检查推定的基因调控元件。与小鼠的类似实验相比,小鸡电穿孔具有快速,简便和廉价的优势。该视频首先演示了如何在蛋壳上开一个窗口来操纵胚胎。接下来,用注入到胚胎下方的印度墨水稀释溶液使胚胎可视化。用玻璃针和移液管将DNA和Fast Green染料注入正在发育的神经管中,然后将铂电极与胚胎平行放置,并用脉冲发生器施加短电脉冲。最后,用胶带将鸡蛋密封,然后放回培养箱中进行进一步发育。此外,视频还显示了正确的卵存储和处理方式,并讨论了电穿孔后胚胎丢失的可能原因。

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