家蚕丝腺分离snRNA-seq和空间转录组学研究方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-01-12 DOI:10.1016/j.xpro.2024.103581

Yan Ma, Keshu Dong, Jie Hu, Yiyun Tang, Hanfu Xu

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引用次数: 0

摘要

家蚕的丝腺（SG）专门合成丝蛋白，因此对丝绸的产量和质量有很大影响。在此，我们介绍了一种从蚕体内分离 SG 细胞核并获得高质量组织切片用于空间转录组学研究的方案。我们介绍了饲养、解剖和细胞核分离的步骤。然后我们详细介绍了包埋、冷冻切片、RNA捕获和测序的步骤。该方案能以单细胞分辨率探索 SG 细胞的空间分布。有关该方案使用和执行的完整细节，请参阅 Ma 等人的文章1。

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Protocol for the isolation of silk glands from silkworms for snRNA-seq and spatial transcriptomics.

The silk glands (SGs) of silkworms specifically synthesize silk proteins, thus strongly influencing the yield and quality of silk. Here, we present a protocol for isolating SG nuclei from silkworms and obtaining high-quality tissue slices for spatial transcriptomics. We describe steps for rearing, dissecting, and nucleus isolation. We then detail procedures for embedding, frozen section, and RNA capturing and sequencing. This protocol enables the exploration of the spatial distribution of SG cells at single-cell resolution. For complete details on the use and execution of this protocol, please refer to Ma et al.¹.

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