AnatomyOrganismsDiseasesChemicals and DrugsAnalytical, Diagnostic and Therapeutic Techniques and EquipmentPhenomena and ProcessesDisciplines and OccupationsInformation Science
Gene Expression RegulationReverse Transcriptase Polymerase Chain ReactionProtein BiosynthesisTranscription, GeneticBlotting, NorthernRNA StabilityCells, CulturedIn Situ HybridizationAmino Acid SequenceBase SequenceCloning, Molecular3' Untranslated RegionsCell LineRNA-Binding ProteinsNucleic Acid HybridizationMolecular Sequence DataRNA, MessengerPoly ADNA, ComplementaryGene Expression Regulation, EnzymologicPolymerase Chain ReactionDNA PrimersRats, Sprague-DawleyLiverPolyribosomesDNABlotting, WesternUp-RegulationPromoter Regions, GeneticRNATranscription FactorsImmunohistochemistryDown-RegulationRNA SplicingTransfectionTumor Cells, CulturedNonsense Mediated mRNA DecayRNA TransportAlternative SplicingTime FactorsDNA-Binding ProteinsCarrier ProteinsMutationCycloheximideTissue DistributionmRNA Cleavage and Polyadenylation FactorsGenesRats, WistarRNA ProbesExonsKineticsDactinomycinSignal TransductionGene Expression Regulation, Developmental5' Untranslated RegionsSequence Homology, Amino AcidGene Expression ProfilingCell NucleusOrgan SpecificityHu Paraneoplastic Encephalomyelitis AntigensGene Expression Regulation, NeoplasticRNA, NeoplasmCell Line, TumorPlasmidsTumor Necrosis Factor-alphaNucleic Acid ConformationNuclear ProteinsCell DifferentiationHeLa CellsMembrane ProteinsRNA, ViralEnzyme InductionIsoenzymesProtein IsoformsRecombinant ProteinsRNA, FungalFibroblastsRibonucleoproteinsReal-Time Polymerase Chain ReactionCytoplasmProtein BindingProteinsDose-Response Relationship, DrugDexamethasoneKidneyPolyadenylationGlobinsInterleukin-1RibosomesRNA, Small InterferingGenes, ReporterIntronsRibonucleasesSequence AlignmentEndoribonucleasesBinding SitesCell DivisionDNA ProbesRestriction MappingOligonucleotide Probes
CDNAProteinAnalysesTissuesVitroProtection AssaysExpressionHybridizationGenomicDownstream applicationsAnalysisIsolationEmbryonicTranscriptsPurificationAmplificationProbesSequenceElectrophoresisSimilarityYeastRadioactiveAbnormalLevelsProcedureComplementaryWesternContrastOligoBeadsIncreasesGene productCellCellsGenesLevelTargetIncreaseSpecificHighSizeTechniqueQuantityHighlyMajor
CDNA5
  • Commonly cDNA is created with labelled primers for the RNA sequence of interest to act as the probe in the northern blot. (wikipedia.org)
  • Take mRNA and use RT to make cDNA labeled with flourescent nucleotides. (flashcardmachine.com)
  • We used the cDNA to develop an RNA (Northern) blot to quantify cellular Hsp90 mRNA levels. (usgs.gov)
  • Northern blot analysis of Jurkat poly(A)+ RNA shows a transcript of 4.7 kb upon probing with the B cell CR2 cDNA. (nih.gov)
  • RNA is ideal for Northern blotting, ribonuclease protection assay, SI nuclease assay, RT-PCR/Q-PCR/RACE analysis, cDNA synthesis, RNA differential display, microRNA studies, and purification of mRNA for library construction. (delos.info)
Protein7
  • E1A-PLECs and E1A-NLFs positive for E1A DNA, mRNA and protein expressed cytokeratin and vimentin but not smooth muscle α-actin. (ersjournals.com)
  • Both exhibited cuboidal morphology and junctional complexes, but did not contain lamellar bodies or express surfactant protein A, B or C mRNAs. (ersjournals.com)
  • Because alternative RNA splicing allows different types of mRNA molecules to be created from a single gene, it generates the diversity of protein function and structure that is essential to complex organisms. (genomicglossaries.com)
  • Highly specific and effective gene silencing of any disease can be achieved by an accurate knowledge of the target mRNA sequence and rational design of its complementary antisense agents for the downregulation of its protein message. (genomicglossaries.com)
  • Despite the proposed role for nerve growth factor (NGF) in this sprouting, we observed no change in NGF mRNA or protein at several postlesion time points. (jneurosci.org)
  • This result was in agreement with the high amount of CYP3A4 protein and mRNA revealed by Western and Northern blot analyses. (aspetjournals.org)
  • The double-stranded RNA duplex leads RNA-induced silencing complex (RISC), containing a member of the conserved Argonaute (Ago) protein family, to target sites of mRNAs, resulting in the destabilization of the mRNAs and/or inhibition of translation [ 4 , 5 ]. (biomedcentral.com)
Analyses4
  • These cells were characterised by PCR, immunocytochemistry, electron microscopy, and Western and Northern blot analyses. (ersjournals.com)
  • The predicted size of the COMT transcript was 1.8-2.0 kb, which could be confirmed from Northern hybridization analyses of the isolated rat liver mRNA. (lu.se)
  • Cells were treated for 3 days with nontoxic concentrations of the drugs, and CYP3A induction was assessed through midazolam hydroxylase activity and Western and Northern blot analyses. (aspetjournals.org)
  • We performed northern blot analyses for the guide RNA truncations and found increased levels of expression for the longer tracrRNA sequences, suggesting that improved target cleavage was at least partially due to higher sgRNA expression or stability ( Fig. 1c ). (cdc.gov)
Tissues4
  • 1) See which mRNAs are being expressed in which tissues. (flashcardmachine.com)
  • The Dynabeads™ mRNA DIRECT™ Kit is designed for simple and rapid isolation of pure, intact polyadenylated (polyA) mRNA directly from the crude lysate of animal and plant cells and tissues. (thermofisher.com)
  • In addition, we studied the mRNA expression of the six mIGFBPs during development and in various adult tissues. (eur.nl)
  • In addition, computer programs usually predict targets on a genome-wide scale irrespective of mRNA and miRNA that are specific to tissues or cell lines. (biomedcentral.com)
Vitro2
  • One polypeptide of 25 kDa, could be immunoprecipitated with anti-COMT antibody from in vitro translation of rat liver mRNA. (lu.se)
  • Northern blots revealed a clear increase in dystroglycan mRNA on day two of in vitro development. (rupress.org)
Protection Assays1
  • In contrast, mRNA levels are now shown not to change by both Northern and RNAse protection assays. (embl.de)
Expression10
  • The northern blot, or RNA blot, is a technique used in molecular biology research to study gene expression by detection of RNA (or isolated mRNA) in a sample. (wikipedia.org)
  • With northern blotting it is possible to observe cellular control over structure and function by determining the particular gene expression rates during differentiation and morphogenesis, as well as in abnormal or diseased conditions. (wikipedia.org)
  • The gene expression of Bax, Bcl-2 and Bcl-xL was analysed by Northern blotting. (portlandpress.com)
  • RNA was extracted from some cells and the extent of expression of inducible NOS mRNA was determined by Northern blotting. (cdc.gov)
  • The level of expression of inducible NOS mRNA was also increased by TDI. (cdc.gov)
  • These findings suggest that upregulated expression of p22phox mRNA and enhanced NADH oxidase activity contribute to the impaired endothelium-dependent vasodilation in OLETF rats. (diabetesjournals.org)
  • We have investigated gelsolin modulation in rat aortic SMC by immunohistochemistry, Western blotting and Northern hybridization using three models which are known to show modulation of actin isoform expression: development, aortic intimal thickening after experimental endothelial injury, and growth in culture. (unige.ch)
  • In liver, a peak of mIGFBP-1 mRNA expression was found around birth, suggesting a function for mIGFBP-1 in the newborn mouse. (eur.nl)
  • The analysis of miRNA targetome from the Ago2 complexes indicated that a lot of mRNAs with high expression level existed in nucleus. (biomedcentral.com)
  • In situ hybridization revealed strong expression of dystroglycan mRNA in all studied epithelial sheets, but not in endothelium or mesenchyme. (rupress.org)
Hybridization3
  • Northern blotting involves the use of electrophoresis to separate RNA samples by size, and detection with a hybridization probe complementary to part of or the entire target sequence. (wikipedia.org)
  • RNase inhibiting agents in the Lysis/Binding Buffer together with stringent hybridization and washing conditions ensure the isolation of pure, intact mRNA from crude samples rich in RNase, without the use of strong chaotropic agents. (thermofisher.com)
  • DIG-labeled hybridization probes are highly sensitive and suitable for low-(single)-copy gene detection of rare mRNA in Southern and northern blots. (thomassci.com)
Genomic2
  • The gene encoding the Ov-phy-1 open reading frame contained 11 introns, similar in structure to the gene encoding human prolyl 4-hydroxylase isoform I. Genomic Southern blot, EST and genomic PCR studies demonstrated that the O. volvulus genome contained between three and eight genes closely related to Ov-phy-1. (embl.de)
  • Amplification products are assayed in genomic Southern blots. (thomassci.com)
Downstream applications1
  • The isolated mRNA is suitable for use in all downstream applications. (thermofisher.com)
Analysis8
  • Northern blot analysis revealed that the Bcl-xL mRNA level increased gradually during the progression towards heart failure. (portlandpress.com)
  • Analysis of Jurkat and Raji mRNA by PCR demonstrated the presence of this novel splice variant in both cell lines. (nih.gov)
  • Northern blot analysis revealed that hOAT3 mRNA is expressed in the kidney, brain, and skeletal muscle. (aspetjournals.org)
  • Primer extension analysis showed that the transcription start point of the rat liver COMT mRNA was 450 bp upstream from the. (lu.se)
  • Employing the DNA blot analysis only one COMT-encoding gene was found in the rat genome. (lu.se)
  • Northern blot analysis showed that the size of the mRNA was about 2 kb, like that of the axi 1 isolated from tobacco. (tubitak.gov.tr)
  • Two cyanate-inducible transcripts of 1500 and 2500nucleotides, respectively, were detected by Northern blot analysis. (embl.de)
  • The target mRNAs in the Argonaute2 (Ago2) complex of nucleus and cytoplasm of gastric cancer cells were analyzed through Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analysis. (biomedcentral.com)
Isolation1
  • The isolation protocol relies on base pairing between the polyA residues at the 3' end of most mRNA, and the oligo (dT) 25 residues covalently coupled to the surface of the Dynabeads. (thermofisher.com)
Embryonic1
  • Chinook salmon embryonic (CHSE-214) cells responded to heat shock with a rapid rise in Hsp90 mRNA through 4 h, followed by a gradual decline over the next 20 h. (usgs.gov)
Transcripts2
Purification1
  • The mRNA purification beads specifically target and capture the mRNA transcriptome from an extremely wide variety of crude starting samples (see protocol). (thermofisher.com)
Amplification2
  • They found that one round of amplification produced approximately 10 3 fold of the estimated amount of starting mRNA, and two rounds produced an approximately 10 5 fold increase. (genomicglossaries.com)
  • High- fidelity mRNA amplification for gene profiling. (genomicglossaries.com)
Probes2
  • Since the gels are fragile and the probes are unable to enter the matrix, the RNA samples, now separated by size, are transferred to a nylon membrane through a capillary or vacuum blotting system. (wikipedia.org)
  • Probes for northern blotting are composed of nucleic acids with a complementary sequence to all or part of the RNA of interest. (wikipedia.org)
Sequence2
  • More specifically, antisense oligonucleotides that are useful as reagents for target validation , or as drugs, are engineered molecules that differ from natural RNA but that have a base sequence that is recognized as being complementary to a very specific mRNA sequence. (genomicglossaries.com)
  • A complementary RNA sequence that binds to a naturally occurring (sense) mRNA molecule, thus blocking its translation. (genomicglossaries.com)
Electrophoresis1
  • Strictly speaking, the term 'northern blot' refers specifically to the capillary transfer of RNA from the electrophoresis gel to the blotting membrane. (wikipedia.org)
Similarity1
  • Northern blotting takes its name from its similarity to the first blotting technique, the Southern blot, named for biologist Edwin Southern. (wikipedia.org)
Yeast1
  • The purpose of this study was to determine the mRNA levels in the yeast mutant rpc128-1007 and its overdose suppressors, RBS1 and PRT1. (mdpi.com)
Radioactive1
  • THEN, transfer to blotting paper and add a radioactive probe to the mRNA you are looking for. (flashcardmachine.com)
Abnormal1
  • With a heterozygote, you would see one normal sized band of the mRNA in question and one abnormal. (flashcardmachine.com)
Levels5
  • The effects of administration of antipsychotic drugs (1-32 days, twice per day) on the rat brain mRNA levels of dopamine D1, D2 and D3 receptors has been assessed by a novel procedure utilising solution hybridisation with oligonucleotides. (nih.gov)
  • Saline and sulpiride (10 mg/kg/injection) had no effect on D1, D2 and D3 receptor mRNA levels. (nih.gov)
  • Haloperidol (1.5 mg/kg/injection) elicited increases in D1, D2 and D3 receptor mRNA levels of 100%, 100% and 300% respectively, after 32 days and loxapine (2 mg/kg/injection) elicited increases of 450%, 150% and 550%, respectively. (nih.gov)
  • Cell density regulates prolyl 4-hydroxylase activity independent of mRNA levels. (embl.de)
  • [ 19 ] studied steady-state levels of the mRNA encoding LEKTI (lymphoepithelial Kazal-type related inhibitor), the gene product of the SPINK5 gene, in cultured epidermal keratinocytes from a healthy control and 5 Netherton syndrome patients. (medscape.com)
Procedure1
  • A general blotting procedure starts with extraction of total RNA from a homogenized tissue sample or from cells. (wikipedia.org)
Complementary2
Western1
  • Western blotting is an important technique used in cell and molecular biology. (delos.info)
Contrast1
  • In contrast, FGF-2 mRNA was increased within 16 hr. (jneurosci.org)
Oligo2
  • Eukaryotic mRNA can then be isolated through the use of oligo (dT) cellulose chromatography to isolate only those RNAs with a poly(A) tail. (wikipedia.org)
  • 1 mg of Dynabeads Oligo (dT) 25 beads (200 µL) binds up to 2 µg of mRNA. (thermofisher.com)
Beads1
Increases1
  • Increases in the amount of PAPP-A mRNA in granulosa cells during follicular development occurs in some but not all species, indicating that other proteases or protease inhibitors may be involved in IGFBP degradation. (bioone.org)
Gene product1
  • To create controls for comparison in a northern blot, samples not displaying the gene product of interest can be used after determination by microarrays or RT-PCR. (wikipedia.org)
Cell4
  • Ok: Use a bead with poly-T to trap the mRNA from a lysed cell (because of the poly-A tail. (flashcardmachine.com)
  • This size is consistent with that of B cell CR2 mRNA. (nih.gov)
  • A typical mammalian cell contains about 10-30 pg of total RNA, from which 1-5% is mRNA. (thermofisher.com)
  • tein, and neuronal cell adhesion molecule mRNA were also increased in olfactory bulb. (cdc.gov)
Cells4
  • Other recent studies verified the presence of PAPP-A mRNA in granulosa cells of humans, monkeys, cattle, mice, and pigs. (bioone.org)
  • Proximal tubule cells exposed to PMA for 2 h demonstrated a two- to threefold increase in Na/H antiporter mRNA (mRNANa/H) abundance 4 h later. (jci.org)
  • In this study, the miRNAs and their target mRNAs in the Argonaute2 (Ago2) complex of nucleus and cytoplasm of gastric cancer cells were characterized using high-throughput sequencing of RNAs isolated by crosslinking immunoprecipitation (HITS-CLIP). (biomedcentral.com)
  • microRNA-mRNA interactions occur in nucleus and cytoplasm of gastric cancer cells. (biomedcentral.com)
Genes3
  • More than half of all human genes are alternatively spliced, so it is expected that alternative pre- mRNA splicing will be responsible for the identification of more than 90 percent of all human proteins. (genomicglossaries.com)
  • Otros autores sugieren que ese es el resultado de mutaciones en oncogenes, genes supresores y enzimas de la vía glucolítica o del metabolismo oxidativo mitocondrial (Myc, Akt, p53, HIF1-α). (researchgate.net)
  • Northern blotting revealed that mIGFBP-2, -3, -4 and -5 genes are already expressed at gestational day 11.5, suggesting a role for these mIGFBPs in embryonal development. (eur.nl)
Level1
  • Hsp90 mRNA level may be useful as a stress indicator, especially in a laboratory setting or in response to acute heat stress. (usgs.gov)
Target1
Increase1
  • This lack of increase contrasts with previous reports as does the mRNA length: this is 50% larger as confirmed by sequencing the 3' end. (embl.de)
Specific1
High2
  • A nylon membrane with a positive charge is the most effective for use in northern blotting since the negatively charged nucleic acids have a high affinity for them. (wikipedia.org)
  • The transfer buffer used for the blotting usually contains formamide because it lowers the annealing temperature of the probe-RNA interaction, thus eliminating the need for high temperatures, which could cause RNA degradation. (wikipedia.org)
Size1
Technique1
  • The northern blot technique was developed in 1977 by James Alwine, David Kemp, and George Stark at Stanford University. (wikipedia.org)
Quantity1
  • The gels can be stained with ethidium bromide (EtBr) and viewed under UV light to observe the quality and quantity of RNA before blotting. (wikipedia.org)
Highly1
  • The mRNA encoding human γ 2 is highly expressed in cerebellum, cerebral cortex, hippocampus and thalamus, whereas γ 3 is abundant in cerebral cortex and amygdala and γ 4 in the basal ganglia. (biomedcentral.com)
Major1
  • The major difference is that RNA, rather than DNA, is analyzed in the northern blot. (wikipedia.org)