Circular RNAs expression profiles in plasma exosomes from early-stage lung adenocarcinoma and the potential biomarkers.
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Circular RNAs expression profiles in plasma exosomes from early-stage lung adenocarcinoma and the potential biomarkers.
This study aims to identify the circular RNA differential (circRNA) in exosomes plasma of patients with lung adenocarcinoma (LUAD) using high-throughput sequencing. First, exosomes were isolated using a kit exosome isolation and confirmed by Western blotting, transmission electron microscopy, and NanoSight Assay. Furthermore, the expression profile circRNA plasma is filtered by high-throughput sequencing and confirmed by fluorescence quantitative real-time polymerase chain reaction (qRT-PCR) and sequencing Sanger.
Finally, network-miRNA-mRNA circRNA performed to predict the potential function circRNAs. The results of the high-throughput sequencing data documented that exosomal circRNAs 182 differentially expressed in all screened, including 105 upregulated and 78 were downregulated in plasma LUAD patients compared with controls. Fourth circRNAs regulated including circ_0001492, circ_0001346, circ_0000690, and circ_0001439 synonymous with sequencing data by qRT-PCR, and the latent interaction circRNA-miRNA-mRNA were exhibited. Taken together, our studies first revealed altered expression exosomal circRNA from plasma samples in patients with LUAD and support the need to explore their potential as biomarkers and pathological effects of lung cancer.
Exosomes are vesicles with sizes ranging from 30-150 nm. Analysis and detection of blood exosomes offer an effective service for cancer diagnosis, prognosis assessment, and evaluation of disease therapies. Because of the difference in separation procedures, methods of collection and use of anticoagulants, serum and plasma samples showed the diversity of the test results.
In order to evaluate the effect of exosomes isolation in serum and plasma samples, two methods commonly used insulation exosomal, ultracentrifugation and precipitation kit based polymers, used, respectively. And the insulating effect was evaluated by comparing the composition and abundant proteins from exosomes isolated based on the analysis of MS-based proteomics. The results showed that plasma exosomes extracted by ultracentrifugation more exosome identify biomarkers and biomarker concentration is higher than others.
And plasma exosomes could be a good example for the blood-based proteomics study of exosomes. It would be useful for future targeted biomarker discovery. This article is protected by copyright. All rights reserved.
Circular RNAs expression profiles in plasma exosomes from early-stage lung adenocarcinoma and the potential biomarkers.
Exosomes molecular diagnostics: direct conversion of exosomes into cDNA for gene amplification by polymerase chain reaction two steps.
Exosomes are cells derived lipid nanoparticles with a size of 30-100 nm in diameter, was found in almost all biological fluids. The composition of exosomes mainly lipids, proteins, RNA, DNA, and non-coding RNA. At present, most of the available methods and commercial kits for exosomal RNA (Exo-RNA) isolation has its limitations and shortcomings.
Starting small volume of exosomes and the use of extraction / filtration column is usually not enough outcome results exosomal RNA after isolation. The majority of RNA contained in exosomes purified various sizes 15-500 nucleotides. Some RNA isolation kit is suitable for the isolation of small RNA transcripts larger but difficult to detect mRNA transcripts. For all kits, gift cost per sample analyzed is very high.
PEG-it Virus Precipitation Solution (100 mL aliquot)
Description: SDS-Blue™ is an innovative patented formula, based on Coomassie blue, that comes in a convenient ready to use format for staining proteins in SDS-PAGE (sodium dodecyl sulphate–polyacrylamide gel electrophoresis). The formulation of SDS-Blue™ provides numerous advantages compared to the classic Coomassie staining or to other similar protein stains. SDS-Blue™ provides higher sensitivity, virtually no background and eliminates the need for destaining of the gel due to its high specificity and affinity to bind to protein only. Not only does SDS-Blue™ yield clear and sharp bands, but it also contains no methanol and acetic acid, making it non-hazardous, safe to handle and friendly to the environment when disposed of. Two other advantages that make SDS-Blue™ the better option is that it is not light sensitive and can be stored at ambient temperature for 24 months. And this provides a considerable convenience, especially to laboratories that need and keep big amount of protein staining solutions – no more jammed refrigerators, you can keep SDS-Blue™ wherever it is most convenient for You!
Thrombin Plasma prep for Exosome precipitation (500 ul at 500U/mL)
Description: A rapid test for detection of antibodies (IgG and IgM) for 2019-nCoV, the novel Coronavirus from the Wuhan strain. The test is easy to perform, takes 10 minutes to provide reliable results and is higly specific to the 2019-nCoV Coronavirus.
Description: A rapid test for detection of antibodies (IgG and IgM) for 2019-nCoV, the novel Coronavirus from the Wuhan strain. The test is easy to perform, takes 10 minutes to provide reliable results and is higly specific to the 2019-nCoV Coronavirus.
Description: Can be used for various proteomics studies in both normal and pathological cases. It is an excellent control and suitable for educational purposes. This product is prepared from whole tissue homogenates and has undergone SDS-PAGE quality control analysis. The protein is stored in a buffer with protease inhibitor cocktail fo prevent degradation.
Description: Can be used for various proteomics studies in both normal and pathological cases. It is an excellent control and suitable for educational purposes. This product is prepared from whole tissue homogenates and has undergone SDS-PAGE quality control analysis. The protein is stored in a buffer with protease inhibitor cocktail fo prevent degradation.
Description: Can be used for various proteomics studies in both normal and pathological cases. It is an excellent control and suitable for educational purposes. This product is prepared from whole tissue homogenates and has undergone SDS-PAGE quality control analysis. The protein is stored in a buffer with protease inhibitor cocktail fo prevent degradation.
Description: A rapid test for detection of antibodies (IgG and IgM) for 2019-nCoV, the novel Coronavirus from the Wuhan strain. The test is easy to perform, takes 10 minutes to provide reliable results and is higly specific to the 2019-nCoV Coronavirus.
Description: The anti-IgY beads were made by cross-linking of bovine IgG anti-IgY antibodies to Protein A/G agarose beads. IgY is the original designation of chicken IgG like immunoglobulin.
Description: Recombinant Human IL-4 produced in E.Coli is a single, non-glycosylated polypeptide chain containing 130 amino acids and having a molecular mass of 15000 Dalton. The rHuIL-4 is purified by proprietary chromatographic techniques.
Description: Recombinant human interleukin-2 is a sterile protein product for injection. rHuIL-2 is produced by recombinant DNA technology using Yeast. It is a highly purified protein containing 133 amino acids, with cysteine mutated to alanine at 125 amino acid position, and has a molecular weight of approximately 15.4kD, non-glycosylated.
Description: Recombinant human interleukin-2 is a sterile protein product for injection. rHuIL-2 is produced by recombinant DNA technology using Yeast. It is a highly purified protein containing 133 amino acids, with cysteine mutated to alanine at 125 amino acid position, and has a molecular weight of approximately 15.4kD, non-glycosylated.
Description: Recombinant human GM-CSF produced in E.coli is a single, non-glycosylated, polypeptide chain containing 127 amino acids, two pairs of disulfide bonds and having a molecular mass of approximately 14.5kD.
Description: Monkeypox virus is the virus that causes the disease monkeypox in both humans and animals. Monkeypox virus is an Orthopoxvirus, a genus of the family Poxviridae that contains other viral species that target mammals. The virus is mainly found in tropical rainforest regions of central and West Africa. The primary route of infection is thought to be contact with the infected animals or their bodily fluids. The genome is not segmented and contains a single molecule of linear double-stranded DNA, 185000 nucleotides long. The Monkeypox Virus real time PCR Kit contains a specific ready-to-use system for the detection of the Monkeypox Virusthrough polymerase chain reaction (PCR) in the real-time PCR system. The master contains reagents and enzymes for the specific amplification of the Monkeypox Virus DNA. Fluorescence is emitted and measured by the real time systems ́ optical unit during the PCR. The detection of amplified Monkeypox Virus DNA fragment is performed in fluorimeter channel 530nm with the fluorescent quencher BHQ1. DNA extraction buffer is available in the kit and serum or lesion exudate samples are used for the extraction of the DNA. In addition, the kit contains a system to identify possible PCR inhibition by measuring the 560nm fluorescence of the internal control (IC). An external positive control defined as 1×10^7 copies/ml is supplied which allow the determination of the gene load.
Description: Monkeypox virus is the virus that causes the disease monkeypox in both humans and animals. Monkeypox virus is an Orthopoxvirus, a genus of the family Poxviridae that contains other viral species that target mammals. The virus is mainly found in tropical rainforest regions of central and West Africa. The primary route of infection is thought to be contact with the infected animals or their bodily fluids.The genome is not segmented and contains a single molecule of linear double-stranded DNA, 185000 nucleotides long.The Monkeypox Virus real time PCR Kit contains a specific ready-to-use system for the detection of the Monkeypox Virusthrough polymerase chain reaction (PCR) in the real-time PCR system. The master contains reagents and enzymes for the specific amplification of theMonkeypox VirusDNA. Fluorescence is emitted and measured by the real time systems ́ optical unit during the PCR. The detection of amplified Monkeypox Virus DNA fragment is performed in fluorimeter channelFAM with the fluorescent quencher BHQ1. DNA extraction buffer is available in the kit and serum or lesion exudate samples are used for the extraction of the DNA. In addition, the kit contains a system to identify possible PCR inhibition by measuring the HEX/VIC/JOE fluorescence of the internal control (IC). An external positive control defined as 1×107copies/ml is supplied which allow the determination of the gene load.
Our current method provides a new way for the direct conversion of exosomes into cDNA synthesis (Exo-cDNA) and subsequent gene detection by the polymerase chain reaction (PCR). This method has several advantages over the established kit available. There is no extraction column is used in this procedure, which means a total recovery of RNA exosomal with maximum results.
