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RNase protection assay on mitochondrial (−)strand transcripts mapping around the cyt b/ND1-binding site in DmTTF-depleted cells

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posted on 31.12.2011, 06:19 by Marina Roberti, Francesco Bruni, Paola Loguercio Polosa, Maria Nicola Gadaleta, Palmiro Cantatore

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Taken from "The termination factor DmTTF regulates mitochondrial transcription"

Nucleic Acids Research 2006;34(7):2109-2116.

Published online 28 Apr 2006

PMCID:PMC1450328.

© The Author 2006. Published by Oxford University Press. All rights reserved

() Schematic representation of digestion products using probes Ribo-1 (295 nt) and Ribo-2 (218 nt). Riboprobes (grey bold arrows), mature transcripts (continuous arrows) and read-through transcripts (dotted arrows) are indicated above the cyt b/ND1 region map. Dashed regions indicate non-coding sequences. () Total cellular RNA (50 µg), extracted from untreated (control) and DmTTF-dsRNA treated (RNAi) D.Mel-2 cells, was hybridized with about 1.5 × 10 c.p.m. of Ribo-1 and Ribo-2 probes and digested with RNase A and T1. Digestion products were denatured and run on a 10% polyacrylamide/7 M urea gel. Y, sample containing 50 µg of yeast total RNA. M, Decade RNA marker (Ambion).

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