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Sample GSM5740380 Query DataSets for GSM5740380
Status Public on Dec 01, 2023
Title Treat_1
Sample type SRA
 
Source name Cal27
Organism Homo sapiens
Characteristics cell line: Cal27
cell type: Human oral cell line
donor: purchased from ATCC
culture time: 6h
treatment: BSArGO@ZIF-8 NSs
Treatment protocol Cells were treated with BSArGO@ZIF8 NSs at a multiplicity of infection (MOI) of F. nucleatum : cell =10:1, and culured at 3d, 7d, 14d and 21d
Growth protocol Cells were cultured with 6-well plates at 37°C in a humidified atmosphere of 5% CO2.
Extracted molecule total RNA
Extraction protocol Total RNA was isolated, evaluated the quality, reverse-transcribed to cDNA,
The total RNA quantity and purity were analysis of Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit (Agilent, CA, USA, 5067-1511) , high-quality RNA samples with RIN number > 7.0 were used to construct sequencing library.After total RNA was extracted, mRNA was purified from total RNA (5ug) using Dynabeads Oligo (dT) (Thermo Fisher, CA, USA) with two rounds of purification. Following purification, the mRNA was fragmented into short fragments using divalent cations under elevated temperature (Magnesium RNA Fragmentation Module (NEB, cat.e6150, USA) under 94℃ 5-7min). Then the cleaved RNA fragments were reverse-transcribed to create the cDNA by SuperScript™ II Reverse Transcriptase (Invitrogen, cat.1896649, USA), which were next used to synthesise U-labeled second-stranded DNAs with E. coli DNA polymerase I (NEB, cat.m0209, USA), RNase H (NEB, cat.m0297, USA) and dUTP Solution (Thermo Fisher, cat.R0133, USA). An A-base was then added to the blunt ends of each strand, preparing them for ligation to the indexed adapters. Each adapter contained a T-base overhang for ligating the adapter to the A-tailed fragmented DNA. Dual-index adapters were ligated to the fragments, and size selection was performed with AMPureXP beads. After the heat-labile UDG enzyme (NEB, cat.m0280, USA) treatment of the U-labeled second-stranded DNAs, the ligated products were amplified with PCR by the following conditions: initial denaturation at 95℃ for 3 min; 8 cycles of denaturation at 98℃ for 15 sec, annealing at 60℃ for 15 sec, and extension at 72℃ for 30 sec; and then final extension at 72℃ for 5 min. The average insert size for the final cDNA librarys were 300±50 bp. At last, we performed the 2×150bp paired-end sequencing (PE150) on an Illumina Novaseq™ 6000 (LC-Bio Technology CO., Ltd., Hangzhou, China) following the vendor's recommended protocol.
 
Library strategy RNA-Seq
Library source transcriptomic
Library selection cDNA
Instrument model Illumina NovaSeq 6000
 
Data processing Reads obtained from the sequencing machines includes raw reads containing adapters or low quality bases which will affect the following assembly and analysis. Thus, to get high quality clean reads, reads were further filtered by Cutadapt (https://cutadapt.readthedocs.io/en/stable/, version:cutadapt-1.9).
Genome_build: hg38
 
Submission date Dec 17, 2021
Last update date Dec 01, 2023
Contact name Wenyan Kang
E-mail(s) 202090000024@sdu.edu.cn
Organization name Shandong university
Street address No. 44-1 Wenhua Road West
City Jinan, Shandong
ZIP/Postal code 250012
Country China
 
Platform ID GPL24676
Series (1)
GSE191172 In situ Growth of ZIF-8 Nanoparticles on Graphene Oxide Nanosheets: A Multifunctional Nanoplatform for Combined Ion‐interference and Photothermal Therapy
Relations
BioSample SAMN24173980
SRA SRX13442791

Supplementary data files not provided
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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