Riboex regent

Manufactured by GeneAll

RiboEX is a reagent used for the extraction and purification of total RNA from various biological samples. It is a single-step, phenol-free method that yields high-quality RNA suitable for downstream applications such as RT-PCR, Northern blotting, and microarray analysis.

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Lab products found in correlation

Improm 2, by Promega (1 mentions) G taq polymerase, by Cosmogenetech (1 mentions) Improm 2 reverse transcriptase, by Promega (1 mentions)

2 protocols using riboex regent

Tau Exon 10 Alternative Splicing Analysis

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Total RNA was extracted using RiboEX regent (GeneAll, Seoul, Korea) according to instructions from the manufacture. Reverse transcription was performed using oligo-dT₁₈ primer and ImProm-II^™ reverse transcriptase (Promega, Madison, WI, USA) to synthesize first-strand cDNA followed by PCR reaction. In the PCR reaction, a primer pair of E8F/E11R was used to detect alternative splicing of endogenous Tau exon 10, a primer set of pcDNAF/E11R was used to detect exon 10 splicing in Tau minigene, the primer sets pcDNAF/E10R and E10F/pcDNAR were used to detect splicing of intron 9 and 10, respectively. The primer sets A1F/A1R and GAPDHF/GAPDHR were used to detect mRNA expression of HnRNPA1 and GAPDH. The primer sequences are listed in Table S1.

Liu Y., Kim D., Choi N., Oh J., Ha J., Zhou J., Zheng X, & Shen H. (2020). hnRNP A1 Regulates Alternative Splicing of Tau Exon 10 by Targeting 3′ Splice Sites. Cells, 9(4), 936.

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Analyzing Splicing Patterns in Cell Lines

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Total RNA was extracted from MDA MB 231, HeLa and plasmid-DNA transfected cells using RiboEX regent (GeneAll) following manufacturer's protocol. 1ug of total RNA was reverse transcribed using oligo dT₁₈ with ImProm-II^™ reverse transcriptase (Promega) following manufacturer's protocol. 1uL of the cDNA was amplified by PCR reaction using G-Taq polymerase (Cosmo Genetech). Different Primers were used to detect splicing: primers for endogenous Ron exon 10-12 [Exon10-for (5’-CCGCTCGAGCGGACCATGTGTGAGAGGCAGCTTCCAG-3’), Exon12-rev (5’-CCGGAATTCCGGTCCTAGCTGCTTCCTCCGCC-3’)], Primers for Ron mini-gene [Exon10-for, pcDNA-rev (5’-CTAGAAGGCACAGTCGAGGCT-3’)], Primers for endogenous intron 11 splicing [Exon10-for and Exon11-rev (5’-ACAGCGCCCAGCCCAATAT-3’)], primers for endogenous intron 12 splicing [Exon11-for (5’-TATATTGGGCTGGGCGCTGTG-3’) and Exon12-rev], primers for detecting exogenous intron 11 splicing [T7-for (5’-TAATACGACTCACTATAGGG-3’) and Exon11-rev], primers for exogenous intron 12 splicing [Exon11-for and pcDNA-rev], primers for GAPDH [GADPH-for (5’-ACCACAGTCCATGCCATCA-3’), GAPDH-rev (5’-TCCACCACCCTGTTGCTGTA-3’)].

Moon H., Cho S., Loh T.J., Oh H.K., Jang H.N., Zhou J., Kwon Y.S., Liao D.J., Jun Y., Eom S., Ghigna C., Biamonti G., Green M.R., Zheng X, & Shen H. (2014). SRSF2 promotes splicing and transcription of exon 11 included isoform in Ron proto-oncogene. Biochimica et biophysica acta, 1839(11), 1132-1140.

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