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mRNAs half-lives determination

ND Ning Dai

Last updated date: Feb 16, 2021 Views: 848 Forks: 0

original An abbreviated version of this protocol was published in eLife in Jul, 2017
IGF2 mRNA binding protein-2 is a tumor promoter that drives cancer proliferation through its client mRNAs IGF2 and HMGA1
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Proliferating cells were treated with 1 uM Actinomycin D (Sigma) for 12 hr to block transcription. After desired timepoints, the total RNAs were extracted (Qiagen RNeasy kit), followed by DNase digestion (eliminating DNA contamination) and cDNA syntheses using Oligo-dT primers (Qiagen) . The concentration of specific mRNAs were quantified by Real-Time RT-PCR using the SYBR Green.

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How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Dai, N(2021). mRNAs half-lives determination. Bio-protocol Preprint. bio-protocol.org/prep846.
  2. Dai, N., Ji, F., Wright, J., Minichiello, L., Sadreyev, R. and Avruch, J.(2017). IGF2 mRNA binding protein-2 is a tumor promoter that drives cancer proliferation through its client mRNAs IGF2 and HMGA1. eLife. DOI: 10.7554/eLife.27155

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