. . . . . "Bioinformatic Analysis Pipeline"@en . "30-read cut-off for untreated samples; exclusion of cross-mapped sequences; parameter dependency <0.3"@en . "Origin 2021b (v9.8.5.212)"@en . "READemption (v0.4.3)"@en . "cutadapt (v2.8)"@en . "segemehl (v0.2.0-418)"@en . "Adapter trimming; read alignment to GRCh38.p13; kinetics model fitting"@en . . . . "CoLoC-seq Experimental Protocol"@en . . . . . . "Differential centrifugation of crude mitochondria"@en . "Organelle Isolation"@en . "14,000 g for 20 min at 4°C to pellet mitochondria"@en . . "Optional isopycnic centrifugation in sucrose gradient for enhanced purity"@en . "Selective sequencing of intact (non-RNase-cleaved) transcripts"@en . "Custom Library Preparation"@en . "3'-adapter ligation to hydroxyl ends"@en . "5'-adapter ligation to phosphorylated ends"@en . "Cap removal (RNA 5'-pyrophosphohydrolase)"@en . "First-strand cDNA synthesis (M-MLV reverse transcriptase)"@en . "PCR amplification (15 cycles, high-fidelity polymerase)"@en . "Size selection: 10-220 nt on agarose gel"@en . . "Excludes RNase-cleaved transcripts (5'-OH and 3'-PO4 incompatible with ligation)"@en . "10 minutes at 25°C"@en . "RNase A Gradient Treatment"@en . "9-10 samples per gradient"@en . "0 - 3 μg/ml RNase A"@en . . "Parallel Mock CoLoC-seq with detergent-permeabilized membranes as control"@en . "High-Throughput Sequencing"@en . "Illumina NextSeq 500"@en . "75 nt"@en . "single-end"@en . . . . . . "The CoLoC-seq experimental protocol integrates organellar isolation, RNase gradient treatment, custom library preparation with selective sequencing of intact transcripts, and kinetics model-based bioinformatic analysis."@en . . "10.1093/nar/gkac1205" . "2023"^^ . . "CoLoC-seq probes the global topology of organelle transcriptomes"@en . . "Nucleic Acids Research"@en . . . . . . . "Tiado Kollender" . "2026-01-11T00:00:00Z"^^ . . . . . . "AIDA-based assertion about the CoLoC-seq experimental protocol"@en . "Nanopublication: CoLoC-seq experimental protocol and methods"@en . . "RSA" . "MIIBIjANBgkqhkiG9w0BAQEFAAOCAQ8AMIIBCgKCAQEAoSR/rqhnkTPQO8u4l2JQAemJo5cN83rT54nA5NXHUqaeu26bEjT/rsX1PCVBhhJ3cV2uu4wjI0NfcVaKPKmODSf6p579tdOykOnjbWLbxxPMe5DC5eV1ThMHsFsNU/6vCYIfS00E7wre7fnFYz7FAG7v1MZEyWeSXsHgk9jHoqQQDPqup5PHReKp1dd6J0Nht/OfSYnUancKAgIFyPhkTlThsLK5xSNyNEgiTR7pwkP5pGq2AnD6bPw4G3kiPnIpIGZNsaeIOvhn64gwbrtoJVdoun/namw6FQ7StEC5km/0evaBZc4qIextwO/s32Wru77ukblJ0Xi/YzbC0iLuIQIDAQAB" . "k+sBTpymBYvAamFSBWMyYKb6xDwo/DvfM1FaHaPytPifKIOcRUDGCk8Fhb8JSRsueOdFCMxcClMCXLUwwKGg+hVfpHGeDKIjrRh2tr4xjsokKJCP5dSg9Tskt/f77++BgRqh02g1B39shCGxXyUHAHIaTwK6aAtjPbJvhVStCm5EGS1LvMKXsrNFLWGHSFBnDeindeFh9GqwbRmd1U1YliQbU+T/ZynxptsZLs64elGrm7Dh4Nfez86L0KE1s+VHM4iUNI0M/5b/441Ir4mr1z4tb8ctiXSLyDkWpD3vQGko4Bdzdq0z6iUZld4i4i3jR5G8j4IYhxISZ+tQGGH2Bg==" . .