PC12 cells were subcultured in six 10-cm culture dishes in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin. The cell culture was performed in an incubator at 37 oC in an atmosphere of 5% CO2 in air and 100% relative humidity. Three of the culture dishes were used as control groups and the rest as test groups. When the cells were grown to 80–90% confluence, LETX-VI was separately added into the three culture dishes of the test groups (final concentration 2.0 µM) to treat the cells for 24 h. All the control and treated cells were collected and total RNA was extracted with a Total RNA Extractor (Trizol) Kit (Sangon Biotech Co., Ltd. Shanghai, China) according to the manufacture’s protocol and used for transcriptome analysis.
Total rna extractor trizol kit
Manufactured by Sangon
Sourced in China
The Total RNA Extractor (Trizol) Kit is a laboratory product designed for the extraction and purification of total RNA from a variety of sample types. The kit utilizes a guanidinium thiocyanate-phenol-chloroform extraction method to isolate RNA.
Automatically generated - may contain errors
Lab products found in correlation
Qubit 2.0 flurometer, by Thermo Fisher Scientific (1 mentions)
Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions)
Cdna synthesis kit, by Illumina (1 mentions)
Ultrasec 2100 pro uv visible spectrophotometer, by Cytiva (1 mentions)
Deep sequencing, by Illumina (1 mentions)
Qubit rna assay kit in qubit 2.0 flurometer, by Thermo Fisher Scientific (1 mentions)
Novaseq sequencer, by Illumina (1 mentions)
Qubit 2.0 fluorometer, by Thermo Fisher Scientific (1 mentions)
Agilent bioanalyzer 2100 system, by Agilent Technologies (1 mentions)
Nanophotometer spectrophotometer, by Implen (1 mentions)
7 protocols using total rna extractor trizol kit
1
LETX-VI Treatment Profiling in PC12 Cells
2
Total RNA Extraction and Sequencing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted using the Total RNA Extractor (Trizol) kit (B511311, Sangon, China) according to the manufacturer’s protocol, and treated with RNase-free DNase I to remove genomic DNA contamination. RNA integrity was evaluated with a 1.0% agarose gel. Thereafter, the quality and quantity of RNA were assessed using a NanoPhotometer ® spectrophotometer (IMPLEN, CA, USA) and a Qubit® 2.0 Flurometer (Invitrogen). The high quality RNA samples were subsequently submitted to the Sangon Biotech (Shanghai) Co., Ltd. for library preparation and sequencing.
3
Total RNA Extraction for RNA-seq
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted using the Total RNA Extractor (Trizol) kit (B511311, Sangon, China) according to the manufacturer’s protocol, and treated with RNase-free DNase I to remove genomic DNA contamination. RNA integrity was evaluated with a 1.0% agarose gel. Thereafter, the quality and quantity of RNA were assessed using a NanoPhotometer® spectrophotometer (IMPLEN, CA, USA) and an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA). The high-quality RNA samples were subsequently submitted to the Sangon Biotech (Shanghai) Co., Ltd. for library preparation and sequencing.
4
Transcriptome Analysis Through RNA Sequencing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted using the Total RNA Extractor (Trizol) Kit (Sangon Biotech Co., Ltd. Shanghai, China). The quality and quantity of total RNA were analyzed using an UltrasecTM 2100 pro UV/Visible Spectrophotometer (Amersham Biosciences, Uppsala, Sweden) and by agarose gel electrophoresis. The cDNA Synthesis Kit (Illumina Inc., San Diego, CA, USA) was used according to the manufacturer's recommendations to prepare cDNA for library construction and Illumina deep sequencing performed by Sangon Biotech Co., Ltd (Shanghai, China). Alignments were performed using the tool package SOAP2 developed for short oligo nucleotide analysis, allowing up to 2 mismatches with reference sequences. Sequenced reads were aligned to human transcript reference sequences from the ENSEMBL database (Homo_sapiens.GRCh37.55.cdna.all.fa) for expression analysis at gene/transcript levels.
5
RNA-Seq Protocol for Differential Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNAs were extracted from tissues and cell samples using Total RNA Extractor (Trizol) Kit (Sangon Biotech) following the manufacturer's instructions for further RNA-Sequencing. The integrity and quality of total RNAs were assessed by 1% (w/v) agarose gel electrophoresis and the 260 nm/280 nm absorbance ratio using a NanoDrop® ND-1000 spectrophotometer. Briefly, mRNA was gathered using magnetic beads with oligo (dT) and then fragmented into short fragments, which were prepared as templates to synthesize the cDNA library. Sequencing of cDNA library was carried out on Illumina Hiseq™ using the paired-end technology. Reads were mapped aligned to the genome assembly Homo sapiens. GRCh38 using HISAT2 and proceeded statistics by RSeQC. The TPM values were calculated for each transcript using StringTie. Differentially expressed genes (DEGs) were evaluated by taking 2-fold changes, p < 0.05 and q < 0.05 as criteria. Further, Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using DAVID bioinformatics resources 6.8 (https://david.ncifcrf.gov/ ).
6
Total RNA Extraction and Sequencing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted using the Total RNA Extractor (Trizol) kit (Sangon, China) according to the manufacturer’s protocol, and treated with RNase-free DNase I to remove genomic DNA contamination. RNA integrity was evaluated with a 1.0% agarose gel. The quality and quantity of RNA were assessed using a Qubit® RNA Assay Kit in Qubit®2.0 Flurometer (Life Technologies, CA). The RNA samples with high quality were subsequently submitted to the Sangon Biotech (Shanghai, China) Co., Ltd. for library preparation and sequencing. A total amount of 2 μg RNA per sample was used as input material for the RNA sample preparations.
7
Total RNA Extraction and Sequencing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted using the Total RNA Extractor (Trizol) Kit (B511311, Sangon, China) according to the manufacturer’s protocol and treated with RNase-free DNase I to remove genomic DNA contamination. Thereafter, the quality and quantity of RNA were assessed using a NanoPhotometer® spectrophotometer (IMPLEN, CA, USA) and a Qubit® 2.0 Fluorometer (Invitrogen). The high-quality RNA samples were subsequently submitted to Sangon Biotech (Shanghai) Co., Ltd. For library preparation and sequencing. Library quality was assessed on an Agilent Bioanalyzer 2100 system. The libraries were then quantified and pooled. Paired-end sequencing of the library was performed on NovaSeq sequencers (Illumina, San Diego, CA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!