Animal Biotechnology Centre, National Dairy Research Institute, Karnal, India.
Methods Mol Biol. 2023;2647:245-258. doi: 10.1007/978-1-0716-3064-8_13.
Cloning by somatic cell nuclear transfer (SCNT) involves the transfer of a somatic nucleus into an enucleated oocyte followed by chemical activation and embryo culture. Further, handmade cloning (HMC) is a simple and efficient SCNT method for large-scale embryo production. HMC does not require micromanipulators for oocyte enucleation and reconstruction since these steps are carried out using a sharp blade controlled by hand under a stereomicroscope. In this chapter, we review the status of HMC in the water buffalo (Bubalus bubalis) and further describe a protocol for the production of buffalo-cloned embryos by HMC and assays to estimate their quality.
体细胞核移植（SCNT）克隆涉及将体细胞核转移到去核卵母细胞中，然后进行化学激活和胚胎培养。此外，手工克隆（HMC）是一种用于大规模胚胎生产的简单有效的 SCNT 方法。HMC 不需要用于卵母细胞去核和重建的微操作器，因为这些步骤是在体视显微镜下使用手动控制的锋利刀片进行的。在本章中，我们回顾了 HMC 在水牛（Bubalus bubalis）中的现状，并进一步描述了通过 HMC 生产水牛克隆胚胎的方案以及评估其质量的方法。
