Complete mitochondrial Physarum polycephalum RNA
SAMN00191251
External Id
The Ohio State University
INSDC center name
2011-01-19T00:00:00Z
INSDC first public
2025-09-22T21:06:21.458Z
INSDC last update
SRS152873
INSDC secondary accession
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
SRS152873
SRA accession
Plasmodial strain M3CVIII (kindly provided by Dr. Mark Adelman) was grown as macroplasmodia at 26*C in semi-defined medium. Macroplasmodia were harvested directly into ice cold BSS [10 mM Tris-HCl (pH 7.5)/ 0.25 M sucrose], with all subsequent steps carried out at 4*C. Cells were lysed in a Waring blender using two 15 second bursts at half maximum speed. The homogenate was filtered prior to pelleting the nuclei and remaining cell debris by centrifugation for 5 min at 700xg. Mitochondria were pelleted by centrifugation for 5 min at 5800xg, resuspended in BSS, layered over Percoll step gradients (densities 1.044-1.095 g/ml in 1 mM Tris-HCl (pH 7.5)/ 0.25 M sucrose), and centrifuged at 47,800xg for 30 seconds. Mitochondrial fractions were collected from each gradient, diluted slowly with 2.5 volumes of 1 mM Tris-HCl (pH 7.5)/ 0.25 M sucrose, and pelleted by centrifugation at 7,600g for 5 min. Total mitochondrial RNA was isolated using TRIzol reagent (Invitrogen) as specified by the manufacturer. Residual DNA was removed by digestion with DNaseI (Roche) in the supplied buffer.
description
Badhamia polycephala
http://purl.obolibrary.org/obo/NCBITaxon_5791
organism
Complete characterization of the edited transcriptome of the mitochondrion of Physarum polycephalum using deep sequencing of RNA
title