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Then, 200 μL of chloroform was added, mixed by vortexing, and allowed to separate into layers. The mixture was centrifuged at 4 °C and 12,000 rpm for 15 min, and the supernatant was transferred to a new 1.5 mL centrifuge tube. An equal volume of isopropanol (Shanghai, China) was ...
Root tips can sense moisture gradients and grow into environments with higher water potential. This process is called root hydrotropism. Here, we report three closely related receptor-like kinases (RLKs) that play critical roles in root hydrotropism: ALT
Early life stress (ELS) is associated with the development of schizophrenia later in life. The hippocampus develops significantly during childhood and is extremely reactive to stress. In rodent models, ELS can induce neuroinflammation, hippocampal neuron
Cells can be removed from the gel using trypsin for downstream analysis (e.g., cytospinning, protein extraction) Cells can be lysed directly in the gel for RNA extraction with Trizol (1 mL Trizol per gel) Gels can be embedded in OCT and cryosectioned for IF. Common problems Special care...
DNA and RNA extraction. DNA extraction and amplicon sequencing Total genomic DNA was extracted from colon biopsies using Qiagen DNeasy Blood and Tissue kit (Qiagen, Valencia, CA) according to the manufacturer’s instructions. The quality and quantity of the DNA were assessed using a Nanodrop ND-...
The incubation was terminated by the addition of Dole's solution (isopropanol-n-heptane-H2SO4, 40:10:1), and unreacted oleate was extracted four times with n-heptane. The remaining oleoyl-CoA in the aqueous phase was quantified by scintillation counting. Quantitative real-time PCR Measurement ...
FUS binds to single- and double-stranded DNA as well as RNA, and participates in the regulation of gene expres- sion, ranging from transcription, and post-transcrip- tional regulation, to mRNA localization and translation [2, 12–14]. FUS directly associates with RNA polymerase II (RNAP II...
events, including infection, malnutrition and genetic abnormalities, can disrupt placental function or, as in preterm birth, can abruptly change the hormonal environment of the developing brain. Such changes might alter early brain development or increase the susceptibility of the immature brain to ...
The RNA was precipitated in isopropanol and washed in 75% ethanol according to the standard TRIzol protocol. The RNA pellet was then re-suspended in RNase-free water (Ambion, Austin, TX) according to the manufacturer's instructions. RNA quantity and integrity were assessed by examining the ...