Although mammalian embryo development depends on critical protein isoforms that arise from embryo-specific nucleic acid modifications, the role of these isoforms is not yet clear. Challenges arise in measuring protein isoforms and nucleic acids from the same single embryos and blastomeres. Here we present a multimodal technique for performing same-embryo nucleic acid and protein isoform profiling (single-embryo nucleic acid and protein profiling immunoblot, or snapBlot). The method integrates protein isoform measurement by fractionation polyacrylamide gel electrophoresis (fPAGE) with off-chip analysis of nucleic acids from the nuclei. Once embryos are harvested and cultured to the desired stage, they are sampled into the snapBlot device and subjected to fPAGE. After fPAGE, ‘gel pallets’ containing nuclei are excised from the snapBlot device for off-chip nucleic acid analyses. fPAGE and nuclei analyses are indexed to each starting sample, yielding same-embryo multimodal measurements. The entire protocol, including processing of samples and data analysis, takes 2–3 d. snapBlot is designed to help reveal the mechanisms by which embryo-specific nucleic acid modifications to both genomic DNA and messenger RNA orchestrate the growth and development of mammalian embryos.

尽管哺乳动物胚胎发育依赖于胚胎特异性核酸修饰产生的关键蛋白质亚型，但这些亚型的作用尚不清楚。测量来自同一单个胚胎和卵裂球的蛋白质亚型和核酸面临着挑战。在这里，我们提出了一种用于进行同胚胎核酸和蛋白质亚型分析（单胚胎核酸和蛋白质分析免疫印迹或 snapBlot）的多模式技术。该方法将通过分级聚丙烯酰胺凝胶电泳 (fPAGE) 进行的蛋白质异构体测量与细胞核核酸的片外分析相结合。一旦胚胎被收获并培养到所需的阶段，它们就会被取样到 snapBlot 装置中并进行 fPAGE。 fPAGE 后，从 snapBlot 装置中切下含有细胞核的“凝胶托盘”，用于芯片外核酸分析。 fPAGE 和细胞核分析对每个起始样品进行索引，产生相同胚胎多模式测量。整个协议，包括样品处理和数据分析，需要 2-3 天。 snapBlot 旨在帮助揭示基因组 DNA 和信使 RNA 的胚胎特异性核酸修饰协调哺乳动物胚胎生长和发育的机制。