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The Ascl2 is a laboratory instrument designed for use in various scientific applications. It serves as a tool for researchers and scientists, providing functionality for their specific needs. The core function of the Ascl2 is to perform its intended tasks within the laboratory environment.

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2 protocols using ascl2

1

Single-Cell RNA Expression Analysis

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Custom Taqman probes were ordered for Nanog, Otx2, Ascl2, and beta-actin (Thermo Fisher Scientific). Single cells were collected directly into 96-well plates by flow cytometry (MoFlo, Beckman Coulter) and reverse transcription was performed as described previously (45 (link)). Cells that had Ct value of beta-actin >14 and no amplification were excluded from the analysis.
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2

qRT-PCR Analysis of Stem Cell Markers

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cDNA was prepared using a Superscript III cDNA-synthesis kit (18080-044, Thermo Scientific). RT–qPCR was performed using TaqMan probes (Life Technologies) and SYBR Green (Thermo Scientific). The mRNA expression of each gene was normalized to the expression of the housekeeping genes Tbp or Hprt1. Relative expression of gene transcripts was analysed using the 2–ΔΔCt method. The RT–PCR data were collected using QuantStudio 12K Flex Software v.1.6 (Applied Biosystems). The following TaqMan probes were used: Olfm4 (Mm01320260_m1, Thermo Scientific), Lgr5 (Mm00438890_m1, Thermo Scientific), Ascl2 (Mm01268891_g1, Thermo Scientific), Tbp (Mm00446973_m1, Thermo Scientific), Prominin-1 (Mm00477115_m1, Thermo Scientific) and Lrig-5 (Mm00456116_m1, Thermo Scientific). Primer sequences for SYBR Green are described in Supplementary Table 5.
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