- Depletion of nuclear LINE1 RNA in mouse ESCs and embryos.
Depletion of nuclear LINE1 RNA in mouse ESCs and embryos.
STAR protocols (2021-08-20)
Michelle Percharde, Chih-Jen Lin, Miguel Ramalho-Santos
PMID34409306
摘要
LINE1 is the most active and abundant family of retrotransposons; it is implicated in a number of pathologies, as well as in early embryo development. We present a protocol to specifically knockdown LINE1 in mouse embryonic stem cells and embryos, including details for the nucleofection and zygote microinjection of LINE antisense oligos, followed by RNA FISH validation. This protocol can be used in development, as well as other cell types where LINE1 is believed to be expressed. For complete information on the use and execution of this protocol, please refer to Percharde et al. (2018).
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产品描述
Sigma-Aldrich
ESGRO®重组小鼠LIF蛋白, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^7 units/ml.
Sigma-Aldrich
硫酸葡聚糖 钠盐 来源于明串珠菌 属, Molecular Biology, average Mw >500,000 (dextran starting material), contains 0.5-2% phosphate buffer
Sigma-Aldrich
透明质酸酶 来源于牛睾丸, Type IV-S, powder, suitable for mouse embryo cell culture, 750-3000 units/mg solid