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Low-viscosity Matrix Suspension Culture for Human Colorectal Epithelial Organoids and Tumoroids

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Extraction of RNA from Recalcitrant Tree Species Paulownia elongata

Authors: Niveditha Ramadoss and Chhandak Basu, date: 07/20/2018, view: 5152, Q&A: 0

[Abstract] Isolation of pure RNA is the basic requisite for most molecular biology work. Plants contain polyphenols and polysaccharides, which can interfere with isolation of pure RNA from them. Especially hardwood tree species like Paulownia elongata have surplus amount of RNA-binding alkaloids, proteins and secondary metabolites that can further ...

RNA Purification from the Unicellular Green Alga, Chromochloris zofingiensis

Authors: Sean D. Gallaher and Melissa S. Roth, date: 04/05/2018, view: 6188, Q&A: 0

[Abstract] Chromochloris zofingiensis is a unicellular green alga that is an emerging model species for studies in fields such as biofuel production, ketocarotenoid biosynthesis and metabolism. The recent availability of a high-quality genome assembly facilitates systems-level analysis, such as RNA-Seq. However, cells of this alga have a tough cell ...

Isolation of Nuclei in Tagged Cell Types (INTACT), RNA Extraction and Ribosomal RNA Degradation to Prepare Material for RNA-Seq

Authors: Mauricio A. Reynoso, Germain C. Pauluzzi, Sean Cabanlit, Joel Velasco, Jérémie Bazin, Roger Deal, Siobhan Brady, Neelima Sinha, Julia Bailey-Serres and Kaisa Kajala, date: 04/05/2018, view: 12785, Q&A: 0

[Abstract] Gene expression is dynamically regulated on many levels, including chromatin accessibility and transcription. In order to study these nuclear regulatory events, we describe our method to purify nuclei with Isolation of Nuclei in TAgged Cell Types (INTACT). As nuclear RNA is low in polyadenylated transcripts and conventional pulldown methods would ...

Total RNA Extraction from Grape Berry Skin for Quantitative Reverse Transcription PCR and Microarray Analysis

Authors: Mami Suzuki and Katsuhiro Shiratake, date: 04/05/2016, view: 7296, Q&A: 0

[Abstract] Extraction of high quality RNA is an essential step for quantitative reverse transcription PCR (qRT-PCR) and microarray analysis. However, it is not easy to extract high quality RNA from fruit materials, which contain high amounts of polysaccharides, lipids and secondary metabolites. Wan and Wilkins (1994) had developed ‘Hot Borate Method’ to ...

Isolation of Polysome-bound mRNA from Rice Solid Tissues Amenable for RT-PCR and Profiling Experiments

Authors: Mehdi Jabnoune, David Secco, Cécile Lecampion, Christophe Robaglia, Qingyao Shu and Yves Poirier, date: 03/05/2015, view: 12999, Q&A: 0

[Abstract] Polysome profile analysis is a frequently performed task in translational control research that not only enables direct monitoring of the efficiency of translation but can easily be extended with a wide range of downstream applications such as Northern and western blotting, genome-wide microarray analysis or qRT-PCR. Here, we describe a method for ...

Polysomal-mRNA Extraction from Arabidopsis by Sucrose-gradient Separation

Authors: Shih-Yun Lin and Guang-Yuh Jauh, date: 12/20/2014, view: 10846, Q&A: 0

[Abstract] mRNAs surrounded by polysomes are ready for translation into proteins (Warner et al., 1963); these mRNAs are defined as polysomal-mRNAs (Mustroph et al., 2009). The process is affected by various growth conditions or surrounding situations. Microarray analysis is a powerful tool for detecting genome-wide gene expression. ...

RNA Extraction from RNase-Rich Senescing Leaf Samples

Author: Susheng Gan, date: 08/20/2012, view: 10638, Q&A: 0

[Abstract] Isolation of intact, full-length high quality RNAs is essential for RNA sequencing, reverse transcription PCR analysis of gene expression as well as RNA gel blot analysis. This simple yet easy protocol is developed to meet this need; in addition to regular samples, this protocol is especially good for isolating RNAs from RNase-rich samples such ...

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