五、送样要求(建议送样量) 1)样品需求:动物样品,总量≥10μg,浓度≥ 150 ng/μl;植物、真菌样品,总量≥20μg,浓度≥ 250 ng/μl。注:目前服务包含的物种有人,老鼠,斑马鱼。其它物种请联系技术评估。 2)样品纯度:动物样品, RIN ≥7,28S/18S≥1.0;植物、真菌样品,RIN ≥6.5, OD260/280≥1.8,28S/18S≥1.0 3)样品请置于1.5 ml 管中,管上注明样品名称、浓度以及制备时间,管口使用Parafilm 封口。在运输前将所有样品管固定于50 ml 带盖离心管,干冰运输。注:RNA 可以提取也可以冻存TRIZOL 中运输,建议冻存TRIZOL 中运输。
六、参考文献 1. 干细胞分化: Lackford B, Yao C, Charles GM, Weng L, Zheng X, Choi EA, et al. Fip1 regulates mRNA alternative polyadenylation to promote stem cell self-renewal. EMBO J 2014;33:878–89. Sommerkamp, P., et al., Differential Alternative Polyadenylation Landscapes Mediate Hematopoietic Stem Cell Activation and Regulate Glutamine Metabolism. Cell Stem Cell, 2020. 2. 癌症: Masamha CP, Xia Z, Yang J, Albrecht TR, Li M, Shyu AB, et al. CFIm25 links alternative polyadenylation to glioblastoma tumour suppression. Nature 2014;510:412–6. Yang, S.W., et al., A Cancer-Specific Ubiquitin Ligase Drives mRNA Alternative Polyadenylation by Ubiquitinating the mRNA 3' End Processing Complex. Mol Cell, 2020. 77(6): p. 1206-1221 e7. 3. 免疫反应: Gruber, A.R., et al., Global 3' UTR shortening has a limited effect on protein abundance in proliferating T cells. Nat Commun, 2014. 5: p. 5465. Jia, X. et al. The role of alternative polyadenylation in the antiviral innate immune response. Nature communications 8, 14605, doi:10.1038/ncomms14605 (2017). Chang, J.W., et al., mRNA 3'-UTR shortening is a molecular signature of mTORC1 activation. Nat Commun, 2015. 6: p. 7218. 4. 神经活动: Oktaba K, Zhang W, Lotz TS, Jun DJ, Lemke SB, Ng SP, et al. ELAV links paused Pol II to alternative polyadenylation in the Drosophila nervous system. Mol Cell 2015;57:341–8. Vallejos Baier, R., J. Picao-Osorio, and C.R. Alonso, Molecular Regulation of Alternative Polyadenylation (APA) within the Drosophila Nervous System. J Mol Biol, 2017. 429(21): p.3290-3300. 5. 胚胎发育 Progressive lengthening of 3' untranslated regions of mRNAs by alternative polyadenylation during mouse embryonic development. Proc Natl Acad Sci U S A, 2009. 106(17): p. 7028-33. 1. Sommerkamp, P., et al., Differential Alternative Polyadenylation Landscapes Mediate Hematopoietic Stem Cell Activation and Regulate Glutamine Metabolism. Cell Stem Cell, 2020. 2. Vallejos Baier, R., J. Picao-Osorio, and C.R. Alonso, Molecular Regulation of Alternative Polyadenylation (APA) within the Drosophila Nervous System. J Mol Biol, 2017. 429(21): p.3290-3300. 3. Zhou, X., et al., Alternative polyadenylation coordinates embryonic development, sexual dimorphism and longitudinal growth in Xenopus tropicalis. Cell Mol Life Sci, 2019. 76(11): p.2185-2198.