characteristics.text: 2024-01-01
characteristics.characteristic: ENA first public
characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: The Francis Crick Institute
characteristics.characteristic: INSDC center name
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS22561510
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-14680:Alpha-RGvssilTRS_8hpi_1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: not collected
characteristics.characteristic: collection date
characteristics.text: Protocols: Lysis of infected cell monolayer in trizol. Cells were incubated with 200 uL Trizol at room temperature for 15 minutes before collection into 2 mL tubes. Trizol extraction using zymo directzol kit per manufacturer's protocol. a region surrounding the N.iORF3 TRS-B sequence was amplified using Platinum SuperFi II DNA Polymerase (Invitrogen) with primers N3geno_FWD and N3geno_REV which contained 5′ Illumina adapter sequences. PCR cycling conditions were 95 ºC for 5 minutes, followed by 20 cycles of 95 ºC for 30 s, 60 ºC for 30 s, and 72 ºC for 20 s, with a final extension of 72 ºC for 5 minutes. Products were cleaned up using Ampure XP beads (Beckman Coulter) at a ratio of 1.8 x. 1 µL of each sample was taken into a PCR reaction with 5 µL of NEB Q5 High-Fidelity DNA Polymerase 2X Master Mix, 1 µL of NXT-IDT Primer Mix 10 µM unique dual index and 3 µL of H2O. PCR cycling conditions were 95 ºC for 3 minutes, followed by 10 cycles of 95 ºC for 30 s, 55 ºC for 15 s, and 72 ºC for 30 s, with a final extension of 72 ºC for 5 minutes. Libraries were cleaned up using Ampure XP beads at a ratio of 0.8 x. Libraries were quantified using Promega Quantifluor reagents, plate reader and Agilent Tapestation.
characteristics.characteristic: description
characteristics.text: not collected
characteristics.characteristic: geographic location (country and/or sea)
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Severe acute respiratory syndrome-related coronavirus
characteristics.characteristic: organism
characteristics.text: Severe acute respiratory syndrome-related coronavirus
characteristics.characteristic: organism
characteristics.text: Severe acute respiratory syndrome-related coronavirus
characteristics.characteristic: scientific_name
characteristics.text: Alpha-RGvssilTRS_8hpi_1
characteristics.characteristic: title