characteristics.text: SAMN41483305
characteristics.characteristic: External Id
characteristics.text: Riken
characteristics.characteristic: INSDC center name
characteristics.text: 2024-05-31T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2024-05-31T04:08:02.518Z
characteristics.characteristic: INSDC last update
characteristics.text: SRS21381422
characteristics.characteristic: INSDC secondary accession
characteristics.text: live
characteristics.characteristic: INSDC status
characteristics.text: Human
characteristics.characteristic: NCBI submission model
characteristics.text: Human.1.0
characteristics.characteristic: NCBI submission package
characteristics.text: SRS21381422
characteristics.characteristic: SRA accession
characteristics.text: not applicable
characteristics.characteristic: age
characteristics.text: RIKEN BRC
characteristics.characteristic: biomaterial_provider
characteristics.text: HeLa/Fucci(SA)2
characteristics.characteristic: cell_line
characteristics.text: not applicable
characteristics.characteristic: collection_date
characteristics.text: Protocols: HeLa/Fucci(SA)2 cells (RCB2867, Riken BRC) were maintained in Dulbecco’s Modified Eagle Medium (Nacalai Tesque) containing 10% fetal bovine serum (gibco) and 1% penicillin-streptomycin (Sigma-Aldrich). We employed a micromanipulation system integrated with an inverted microscope, featuring two micromanipulators: PatchMan NP2 (Eppendorf) and TransferMan 4r (Eppendorf) to position a glass capillary (HUMANIX, CT-0P) proximity to a cellular membrane. One micromanipulator was equipped with a capillary holder (Pipette holder, Narishige, H-7), positioned at a 75° angle. The glass capillary was filled with 1 μL of sucrose solution (260 mM Sucrose and 20 mM HEPES) at the tip as an inner solution. A platinum wire of 200-μm in diameter (Niraco, PT-351265) was immersed in the sucrose solution. The another manipulator was equipped with a platinum electrode of 1 mm in diameter (Niraco, PT-351481). The platinum electrode immersed in a sucrose mixture medium approximately 5 mm away from the microscope's field of view. Each platinum wire was linked to a source meter (Keithley, 2410-C) for real-time monitoring of current values and application of the voltage via an RS-232 C serial communication. To detect the contacting of a tip of the glass capillary to the cellular membrane, a constant voltage of 0.3 V was maintained before the sampling. The contact was detected by damping of the ionic current. Upon contact, intracellular products were collected into the capillary using square pulse conditions of 2 Hz, 2 V spacing, -10 V pulse, and 10% duty ratio for 5 s for HeLa/Fucci cells. Subsequently, the capillary was swiftly withdrawn from the medium and detached from the capillary holder to collect the extracted RNA.
characteristics.characteristic: description
characteristics.text: cervical carcinoma
characteristics.characteristic: disease
characteristics.text: not applicable
characteristics.characteristic: geo_loc_name
characteristics.text: subcellular mRNA from single HeLa/Fucci cells obtained by nanoelectrokinetic aspiration
characteristics.characteristic: isolate
characteristics.text: Homo sapiens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: organism
characteristics.text: RNA
characteristics.characteristic: sample_type
characteristics.text: female
characteristics.characteristic: sex
characteristics.text: 2
characteristics.characteristic: source_material_id
characteristics.text: cervix
characteristics.characteristic: tissue
characteristics.text: HeLaFucci, live-cell RNA-seq
characteristics.characteristic: title