Total RNA of tumor samples was isolated using RNeasy Plus Universal Kits (Qiagen, GER). RNA concentration was quantified using QubitTM RNA HS Assay Kit (ThermoFisher Scientific, USA). RNA purity and integrity were analyzed using Take3 (BioTek, USA) and the RNA Cartridge kit of the Qseq100 Bio-Fragment Analyzer (Bioptic, CHN), respectively. Then, RNA-seq libraries were constructed using VAHTS mRNA-seq V3 Library Prep Kit for Illumina (Vazyme, CHN). Libraries were sequenced on the NextSeq 550AR platform with 150 bp paired-end reads. Quality control of WES data was described in Supplementary Data 3 .
Qseq100 bio fragment analyzer
Manufactured by Bioptic
Sourced in China, United States
The Qseq100 Bio-Fragment Analyzer is a lab equipment designed to analyze and quantify DNA or RNA fragments. It utilizes advanced optical detection technology to accurately measure the size and concentration of biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Rna cartridge kit, by Bioptic (3 mentions)
Rneasy plus universal kit, by Qiagen (3 mentions)
Take3, by Agilent Technologies (2 mentions)
Qubit rna hs assay kit, by Thermo Fisher Scientific (2 mentions)
Generead rrna depletion kit, by Qiagen (1 mentions)
Qubit 3.0 spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Exorneasy serum plasma maxi kit, by Qiagen (1 mentions)
Hiseq x ten system, by Illumina (1 mentions)
Qubit 3, by Thermo Fisher Scientific (1 mentions)
Nextseq 550ar, by Illumina (1 mentions)
Vahts mrna seq v3 library prep kit, by Vazyme (1 mentions)
5 protocols using qseq100 bio fragment analyzer
1
Tumor RNA Isolation and Sequencing
2
RNA Extraction and RNA-seq Library Preparation
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Total RNA of tumor samples was isolated from the tumor tissue with RNeasy Plus Universal Kits (Qiagen, GER) according to the manufacturer's instructions. A Qubit™ RNA HS Assay Kit (Thermo Fisher Scientific, USA) was used to quantify the concentrations of the extracted RNA. The purity and integrity of the RNA were checked with Take 3 (BioTek, USA) and the RNA Cartridge kit of the Qseq100 Bio-Fragment Analyzer (Bioptic, CHN), respectively. The VAHTS mRNA-seq V3 Library Prep Kit for Illumina (Vazyme, CHN) was used to construct RNA-seq libraries. Finally, the libraries were sequenced on the NextSeq 550AR with 150 bp paired-end reads.
3
RNA Sequencing Library Preparation
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The total RNA of cells was isolated by using TRIzol and used for RNA sequencing. RNA quantification was performed with a Qubit 3.0 spectrophotometer (Thermo Fisher, MA, USA). Library preparation was performed by Epibiotek (Guangzhou, China). Briefly, total RNA was treated with the GeneRead™ rRNA Depletion Kit (Qiagen, Hilden, Germany, Cat No. 180211) to remove ribosomal RNA. rRNA-depleted RNA was fragmented and then used to construct strand-specific RNA libraries by using the VAHTS Stranded RNA-seq Library Prep Kit for Illumina (Vazyme, Nanjing, China, Cat. No NR602) according to the manufacturer’s instructions. Library quality was determined on a Qseq100 Bio-Fragment Analyzer (Bioptic, Taiwan, China). The strand-specific libraries were sequenced.
4
Exosomal RNA Sequencing Protocol
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The total exosomal RNA was extracted using the exoRNeasy Serum/Plasma Maxi Kit (Qiagen GmbH) according to the manufacturer's protocol and used for mRNA sequencing. RNA quantification was performed with Qubit 3.0 (Thermo Fisher Scientific). For mRNA-seq, library preparation was performed using the Epi™ longRNA Ampli kit (Epibiotek, Guangzhou, China) according to the manufacturer's instructions. In brief, the ribosomal RNA (rRNA) was depleted and the RNA samples were mixed with the provided RT Primer mix and deoxynucleoside triphosphate (dNTP) mix to perform RT and synthesize the first-strand cDNA by PCR. After RT, the second-strand cDNA was synthesized by PCR under optimized reaction conditions. After amplification by PCR, cDNA was purified and size-selected with magnetic beads. Library evaluation and quantification were performed on a Qseq100 Bio-Fragment Analyzer (BiOptic, Taiwan, China), and subsequent next-generation sequencing (NGS) was performed by using an Illumina HiSeq X Ten System (Illumina, San Diego, CA, USA).
5
RNA-seq of Tumor Tissue Samples
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We isolated the total RNA of tumor samples from the tumor tissues via RNeasy Plus Universal Kit (Qiagen, USA). Utilizing the QubitTM RNA HS Assay Kit, the concentrations of the extracted RNA were then quantified. The integrity and purity of the extracted RNA were tested by the RNA Cartridge kit of the Qseq100 Bio-Fragment Analyzer (Bioptic, China) and the Take3 kits (BioTek, USA), respectively. We then generated the RNA-seq libraries by the VAHTS mRNA-seq V3 Library Prep Kit (Vazyme, China). Eventually, the libraries were sequenced on the NextSeq 550AR (Illumina, USA) with 150-bp paired-end reads.
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