Abstract：
The tarsal plate is an eyelid tissue that maintains lid structure from inside the upper/lower eyelids, and it surrounds the meibomian glands and supports their unique secretion mechanism. Sebaceous carcinoma, a malignant eyelid tumour, can sometimes develop from the meibomian glands and is usually excised together with the tarsal plate during surgery, so the tarsal plate serves as a control research tissue. However, since the plate is thick, hard and heterogeneous with few cells, obtaining enough genomic DNA and/or total RNA is often difficult. Therefore, we attempted to establish an efficient protocol to obtain DNA and RNA simultaneously by comparing the combinations of homogenization (mortar/pestle, pellet pestle or SK mill) and purification (organic solvent or spin column) methods using rabbit tarsal plates. Based on the yield, quality and hands-on time, the SK mill and spin column was found to be the most efficient combination. We then applied the established protocol to extract DNA/RNA from six human tarsal-plate samples and succeeded in generating high-quality exome and transcriptome datasets via a next-generation sequencer with sufficient coverage and meibomian gland-specific expression of representative genes, respectively. Our new findings will provide ideal reference data for future genetic and gene-expression studies of sebaceous carcinoma.
摘要：
睑板是眼睑组织，从上/下眼睑内部保持眼睑结构，它围绕睑板腺并支持其独特的分泌机制。皮脂腺癌,恶性眼睑肿瘤,有时可以从睑板腺发展，通常在手术过程中与tar板一起切除，因此，板作为对照研究组织。然而,因为盘子很厚，坚硬且异质，细胞少，获得足够的基因组DNA和/或总RNA通常是困难的。因此,我们试图建立一个有效的方案，通过比较均化的组合来同时获得DNA和RNA（研钵/研钵，球棒或SK磨机）和使用兔tarsal板的纯化（有机溶剂或旋转柱）方法。根据产量，质量和动手时间，SK磨机和旋转柱被发现是最有效的组合。然后，我们应用已建立的协议从六个人tar板样品中提取DNA/RNA，并通过下一代测序仪成功生成高质量的外显子组和转录组数据集，该测序仪具有足够的覆盖率和睑板腺特异性表达的代表性基因，分别。我们的新发现将为皮脂腺癌的未来遗传和基因表达研究提供理想的参考数据。
