Name: analysis of single-cell gene transcription by rna fluorescent in situ hybridization (fish)
Uploaded: 2012-10-07T14:00:00
Description: Watch this Scientific Journal Video about Analysis of Single-cell Gene Transcription by RNA Fluorescent In Situ Hybridization FISH at JoVE.com

Forfatterne vil gerne takke Michael Alifrangis og Ulla Abildtrup til genotypning af parasitter og Christina Holm for fremragende teknisk bistand. Dette arbejde blev finansieret af Howard Hughes Medical Institute (tilskud 55.005.511), Lundbeckfonden (tilskud R9-A840) og af Niels Bohr Instituttet.

1. Generering af Frisk Udvalgte Inficerede Erythrocytter Til dette assay, er de bedste resultater opnået ved anvendelse af frisk udvalgte kulturer for overfladeekspression af PfEMP1 protein. I dette særlige forsøg blev 3D7 P. falciparum linie blev valgt under anvendelse af specifikke antistoffer som beskrevet tidligere 1. Dag 1 <ol><li> Høst 200 pi pakkede blodlegemer fra en P. falciparum kultur indeholdende 2-5% sent IE ved centri...

<ol>
	<li>Joergensen, L., Bengtsson, D. C., Bengtsson, A., Ronander, E., Berger, S. S., Turner, L., Dalgaard, M. B., Cham, G. K., Victor, M. E., Lavstsen, T., Theander, T. G., Arnot, D. E., Jensen, A. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surface+co-expression+of+two+different+PfEMP1+antigens+on+single+Plasmodium+falciparum-infected+erythrocytes+facilitates+binding+to+ICAM1+and+PECAM1.">Surface co-expression of two different PfEMP1 antigens on single Plasmodium falciparum-infected erythrocytes facilitates binding to ICAM1 and PECAM1.</a> PLoS Pathog. 2, 1001083 (2010).</li><li>Pinaud, R., Mello, C. V., Velho, T. A., Wynne, R. D., Tremere, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+two+mRNA+species+at+single-cell+resolution+by+double-fluorescence+in+situ+hybridization.">Detection of two mRNA species at single-cell resolution by double-fluorescence in situ hybridization.</a> Nat. Protoc. 3, 1370-1379 (2008).</li><li>Cheeseborough, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> District Laboratory Practice in Tropical Countries. , (2006).</li><li>Brolin, K. J., Ribacke, U., Nilsson, S., Ankarklev, J., Moll, K., Wahlgren, M., Chen, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+transcription+of+duplicated+var2csa+gene+copies+in+individual+Plasmodium+falciparum+parasites.">Simultaneous transcription of duplicated var2csa gene copies in individual Plasmodium falciparum parasites.</a> Genome Biol. 10, R117 (2009).</li><li>Epp, C., Li, F., Howitt, C. A., Chookajorn, T., Deitsch, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chromatin+associated+sense+and+antisense+noncoding+RNAs+are+transcribed+from+the+var+gene+family+of+virulence+genes+of+the+malaria+parasite+Plasmodium+falciparum.">Chromatin associated sense and antisense noncoding RNAs are transcribed from the var gene family of virulence genes of the malaria parasite Plasmodium falciparum.</a> RNA. 15, 116-127 (2009).</li><li>Freitas-Junior, L. H., Hernandez-Rivas, R., Ralph, S. A., Montiel-Condado, D., Ruvalcaba-Salazar, O. K., Rojas-Meza, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Telomeric+heterochromatin+propagation+and+histone+acetylation+control+mutually+exclusive+expression+of+antigenic+variation+genes+in+malaria+parasites.">Telomeric heterochromatin propagation and histone acetylation control mutually exclusive expression of antigenic variation genes in malaria parasites.</a> Cell. 121, 25-36 (2005).</li><li>Obernosterer, G., Martinez, J., Alenius, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Locked+nucleic+acid-based+in+situ+detection+of+microRNAs+in+mouse+tissue+sections.">Locked nucleic acid-based in situ detection of microRNAs in mouse tissue sections.</a> Nature Protoc. 2, 1508-1514 (2007).</li><li>Li, F., Sonbuchner, L., Kyes, S. A., Epp, C., Deitsch, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nuclear+non-coding+RNA+are+transcribed+from+the+centromeres+of+Plasmodium+falciparum+and+are+associated+with+centromeric+chromatin.">Nuclear non-coding RNA are transcribed from the centromeres of Plasmodium falciparum and are associated with centromeric chromatin.</a> J. Biol. Chem. 283, 5692-5698 (2008).</li><li>Thompson, J., Sinden, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+situ+detection+of+Pbs21+mRNA+during+sexual+development+of+Plasmodium+berghei.">In situ detection of Pbs21 mRNA during sexual development of Plasmodium berghei.</a> Mol. Biochem. Parasitol. 68, 189-196 (1994).</li><li>Thompson, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+situ+detection+of+RNA+in+blood-+and+mosquito-stage+malaria+parasites.">In situ detection of RNA in blood- and mosquito-stage malaria parasites.</a> Methods Mole Med. 72, 225-235 (2002).</li><li>Arnot, D. E., Ronander, E., Bengtsson, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+progression+of+the+intra-erythrocytic+cell+cycle+of+Plasmodium+falciparum+and+the+role+of+the+centriolar+plaques+in+asynchronous+mitotic+division+during+schizogony.">The progression of the intra-erythrocytic cell cycle of Plasmodium falciparum and the role of the centriolar plaques in asynchronous mitotic division during schizogony.</a> Int. J. Parasitol. 41, 71-80 (2011).</li><li>Osborne, C. S., Chakalova, L., Brown, K. E., Carter, D., Horton, A., Debrand, E., Goyenechea, B., Mitchell, J. A., Lopes, S., Reik, W., Fraser, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Active+genes+dynamically+colocalize+to+shared+sites+of+ongoing+transcription.">Active genes dynamically colocalize to shared sites of ongoing transcription.</a> Nat. Genet. 36, 1065-1071 (2004).</li></ol>

RNA FISH-analyse, i modsætning til fremgangsmåder såsom Northern blotting og RT-PCR, muliggør diskriminering af specifikke mRNA-transkripter på enkeltcelleniveau. Dette gør det muligt at skelne mellem transkriptionelt aktive og inaktive celler, i dette eksempel, P. falciparum parasitiske protozoer inden i humane røde blodlegemer. Sådanne helcelle-observationer er ofte nødvendige, og kan udrede vigtige og hidtil ukendte transkriptionelle mønstre en. Selvom andre ...

<table border="1"><tbody><tr><td> Navn på reagenset </td><td> Firma </td><td> Katalognummer </td><td> Kommentarer </td></tr><tr><td> Eddikesyre </td><td> Sigma / Aldrich </td><td> 338.826-100ml </td><td></td></tr><tr><td> Albumax medier: RPMI 1640 Glutamin løsning </td><td> Lonza </td><td> BE12-115F </td><td> 500 ml RPMI 1640 5 ml glutamin opløsning </td></tr><tr><td> Albumax medier: Gentamycin sulfat Albumax-løsning </td><td> Lonza </td><td> BE02-012E </td><td> 2,5 ml gentamycin-sulfat 50 ml Albumax-opløsning </td></tr><tr><td> Albumax-løsning: hypoxanthin </td><td> Sigma-Aldrich </td><td> H9377 </td><td> 0,8 g Hypoxanthin </td></tr><tr><td> Albumax-løsning: AlbuMAX II </td><td> Jegnvitrogen </td><td> 11021-037 </td><td> 200 g Albumax </td></tr><tr><td> Albumax-løsning: RPMI 1640 </td><td> Lonza </td><td> BE12-115F </td><td> 4 liter RPMI 1640 Opløs med magnet på max. 50 ° C. Filtrere steriliseres og opbevares ved -20 ° C i alikvoter. </td></tr><tr><td> Amberlite </td><td> Sigma </td><td> A5710-110G </td><td> Harpiks deionize formamid. </td></tr><tr><td> Anti-biotin ged pAb peroxidasekonjugat </td><td> Calbiochem </td><td> 203.206 </td><td></td></tr><tr><td> Anti-Dig-antistof </td><td> Novus Biologicals </td><td> NB100-41.330 </td><td></td></tr><tr><td> Anti-fade reagens med DAPI </td><td> Invitrogen </td><td> P36931 </td><td> De monteringsmedium skal hærde i 24 timer før forsegling slæden helt. </td></tr><tr><td> Biotin RNA Labeling Mix </td><td&gt; Roche </td><td> 11685597910 </td><td></td></tr><tr><td> Kamera digital </td><td> Nikon digital syn DC-F11 </td><td></td><td></td></tr><tr><td> Dækglas </td><td> Menzel-Glaser </td><td> 631-1570 </td><td></td></tr><tr><td> dyrkningskolbe 25 cm2 Nunclon overflade </td><td> Nunc </td><td> 156.340 </td><td></td></tr><tr><td> DEPC </td><td> Fluka / Sigma </td><td> 32.490-100ml </td><td> 1 ml DEPC i 1000 ml deinonized vand. Tilføj en omrører og omrøres i 12 timer. Autoklav i 30 min. </td></tr><tr><td> Digital kamera </td><td> Nikon digital syn DC-F11 </td><td></td><td></td></tr><tr><td> Dynabeads Protein A </td><td> Invitrogen </td><td> 10002D </td><td></td></tr><tr><td> DynaMaq-15 Magnet </td><td> Invitrogen </td><td> 123-01D </td><td></td></tr><tr><td> Formamid bioultra 99% </td><td> Fluka/ Sigma </td><td> 47671-1l-F </td><td> Deioniseret formamid: 5 g ionbytterharpiks per 100 ml formamid. Der omrøres 30 min. Der filtreres gennem Whatman papir. </td></tr><tr><td> . Gelatine 0 75% opløsning: Gelatine </td><td> Sigma-Aldrich </td><td> G2500 </td><td> 3,75 g gelatine i 500 ml RPMI 1640. Der opvarmes til 56 ° C til opløsning. Filtrere sterilisere ved 56 ° C. Opbevar ved -20 ° C i alikvoter. </td></tr><tr><td> . Gelatine 0 75% opløsning: RPMI 1640 </td><td> Lonza </td><td> BE12-115F </td><td> 3,75 g gelatine i 500 ml RPMI 1640. Der opvarmes til 56 ° C til opløsning. Filtrere sterilisere ved 56 ° C. Opbevar ved -20 ° C i alikvoter. </td></tr><tr><td> Glutamin løsning: L-glutamin </td><td> Sigma-Aldrich </td><td> G3126 </td><td> 14,6 g L glutamin i 500 ml 0,9% NaCl. Opløs, filter sterilize og opbevares ved -20 ° C i alikvoter. </td></tr><tr><td> Glutamin løsning: HCl </td><td> Sigma </td><td> H1758 </td><td> 14,6 g L glutamin i 500 ml 0,9% NaCl. Opløs, filtrer steriliseres og opbevares ved -20 ° C i alikvoter. </td></tr><tr><td> Hybridiseringsopløsning: Formamid Bio ultra 99% SSC 20x </td><td> Fluka / Sigma </td><td> 47671-1l-F </td><td> I alt 20 ml, opbevare frosset ved -20 ° C i alikvoter 10 ml deioniseret formamid </td></tr><tr><td> Hybridiseringsopløsning: Denhardts 50x koncentrat </td><td> Sigma </td><td> D2532 </td><td> 5ml 20xSSC </td></tr><tr><td> Hybridiseringsopløsning: Gær tRNARoche blokeringsreagens </td><td> Sigma </td><td> R-6750 </td><td> 2 ml 50x Denhardts 250 ul 20 mg per ml gær-tRNA </td></tr><tr><td><sTrong&gt; hybridiseringsopløsning: laksesperm DNA </td><td> Fluka / Sigma </td><td> 31.149-106GF </td><td> 0,4 g Roche blokeringsreagens 1 ml 10 mg pr ml laksesperm-DNA (Critical: denaturerer laks spermDNA ved 96 ° C i 5 min før tilsætning til hybridiseringsopløsningen) </td></tr><tr><td> Hybridizer ThermoStar 100 HC4 </td><td> Quantifoil Instruments GmbH </td><td> 1004-0011 </td><td> Kan erstattes af hybridisering ovn og RNase fri hybridiseringsmetoder kamre polstret med DEPC vand. </td></tr><tr><td> Immersionsolie UV transparent fluorescens free </td><td> Sigma </td><td> 10976-1EA </td><td></td></tr><tr><td> Immunofluorescens eller konfokalt mikroskop </td><td> Nikon D-Eclipse TE2000C </td><td></td><td></td></tr><tr><td> Neglelak </td><td> Fås i ethvert apotek </td><td></td><td></td></tr><tr><td> PBS 20x: NaCl KCl Na 2 HPO 4 x 2H 2 O KH 2 PO 4 DEPC deioniseret H 2 O </td><td></td><td></td><td> Ajust pH til 7,4 160 g 4 g 23 g 4 g 1 l </td></tr><tr><td> Paraformaldehyd 4% </td><td> Fluka / Chemika </td><td> 76.240 </td><td> 4 g PFA i 80 ml PBS / DEPC. Der opvarmes til 65 ° C, indtil PFA opløses. Tilføj 20 ml PBS, lad opløsningen køle af. PH justeres til 7,4. Filter. Opbevares i alikvoter ved -20 ° C. </td></tr><tr><td> Paraformaldehyd 4% / eddikesyre 5% </td><td></td><td></td><td> 950 pi paraformaldhyde + 50 ul Eddikesyre </td></tr><tr><td> Pepsin </td><td> Sigma / Aldrich </td><td> P7000 </td><td></td></tr><tr><td> Peroxidasekonjugeret anti-biotin </td><td> Calbiochem </td><td> 203.206 </td><td></td></tr><tr><td> RBC-vask medier: RPMI 1640 Glutamin opløsning </td><td> Lonza </td><td> BE12-115F </td><td> 500 ml RPMI 1640 5 ml glutamin opløsning </td></tr><tr><td> RBC-vask medier: gentamycinsulfat </td><td> Lonza </td><td> BE02-012E </td><td> 2,5 ml gentamycin-sulfat </td></tr><tr><td> RNase </td><td> Sigma / Aldrich </td><td></td><td> En 10 mg / ml stamopløsning. </td></tr><tr><td> RNase free 1,5 ml rør </td><td> Ambion </td><td> AM12450 </td><td></td></tr><tr><td> RNase Zap </td><td> Ambion </td><td> AM9780 </td><td></td></tr><tr><td> Slides 4 brønde 11mm </td><td> Thermo Scientific </td><td> MENZXER306W </td><td></td></tr><tr><td> SSC 20x: NaCl </td><td> Sigma / Aldrich </td><td> S9625 </td><td> 3 M NaCl (175 g / l) 0,3 M Na3-citrat x H2O (88 g / l) indstilles til pH 7,0 med 1 M HCI </td></tr><tr><td> SSC 20x: Natriumcitrat dehydrat </td><td> Sigma / Aldrich </td><td> W302600 </td><td> 3 M NaCl (175 g / l) 0,3 M Na3-citrat x H2O (88 g / l) indstilles til pH 7,0 med 1 M HCI </td></tr><tr><td> SSPE 20x: NaCl </td><td> Sigma / Aldrich </td><td> S9625 </td><td> 175,3 g NaCl </td></tr><tr><td> SSPE 20x: NaH 2 PO 4 </td><td> Sigma / Aldrich </td><td> S0751 </td><td> 27,6 g NaH 2 PO4. </td></tr><tr><td> SSPE 20x: 4 EDTA pulver </td><td></td><td></td><td> 9,4 g EDTA pulver Tilsættes DEPC vand, justere pH 7,4. AutoKlaven i 20 min </td></tr><tr><td> TNB-puffer: TNT-puffer </td><td></td><td></td><td> 10 ml TNT-puffer </td></tr><tr><td> TNB-puffer: Blocking reagent </td><td></td><td></td><td> 0,05 g blokeringsreagens At opløse blokerende reagens, opvarme opløsningen til 60 ° C i en time under omrøring. Opbevar ved -20 ° C. (Afledt af Perkin Elmer TSA-protokollen.) </td></tr><tr><td> TNT buffer: Tris / HCl </td><td> Sigma </td><td> T1503 </td><td> 1M Tris / HCI, pH 8,0 </td></tr><tr><td> TNT buffer: NaCl </td><td> Sigma Aldrich </td><td> S9625 </td><td> 100 ml </td></tr><tr><td> TNT buffer: Tween20 </td><td> Sigma </td><td> 93.773 </td><td> 5 M NaCI 30 ml 1 ml DEPC dH2O869 Ml PH justeres til 7,5 ved stuetemperatur. (Afledt af Perkin Elmer TSA-protokollen.) </td></tr><tr><td> TSA plus Cyanine3 / Fluorescein-system </td><td> Perkin Elmer </td><td> NEL753000IKT </td><td> Læs protokollen fra TSA Plus Fluorescence Palette System omhyggeligt, før du starter eksperimentet. </td></tr><tr><td> Rør 14 ml sterilt </td><td> Almeco - CM LAB Aps </td><td> 91.016 </td><td></td></tr></tbody></table>

analysis of single-cell gene transcription by rna fluorescent in situ hybridization (fish)

Adhæsion af Plasmodium falciparum inficerede erythrocytter (IE) til humane endotel receptorer under malariainfektioner medieres ved ekspression af PfEMP1 proteinvarianter kodet af VaR gener. The haploid P. falciparum genom havne cirka 60 forskellige was gener, hvoraf kun én er blevet menes at blive transskriberet per celle ad gangen i blodet fase af infektionen. Hvordan en sådan gensidigt udelukker regulering af var gentranskription opnås er uklar, da er at identificere individuelle was gener eller undergrupper af VaR gener forbundet med forskellige receptorer og konsekvensen af forskellen binding på det kliniske resultat af P. falciparum infektioner. Senest har gensidigt udelukker transkription paradigme været draget i tvivl ved transskription assays baseret på individuel P. falciparum udskrift identifikation i en enkelt infected erytrocytiske celler under anvendelse af RNA fluorescerende in situ hybridisering (FISH) analyse af var gentranskription af parasitten i enkelte kerner af P. falciparum IE 1. Her præsenterer vi en detaljeret protokol til udførelse af RNA-FISH metode til analyse af var gentranskription i single-kerner af P. falciparum inficerede humane erythrocytter. Fremgangsmåden er baseret på anvendelse af digoxigenin-og biotin-mærket antisense RNA-prober ved hjælp af TSA Plus Fluorescence Palette System 2 (Perkin Elmer), mikroskopiske analyser og frisk valgt P. falciparum IE. Den in situ-hybridisering metode kan bruges til at overvåge transkription og regulering af en række gener udtrykt under de forskellige faser af P. falciparum livscyklus og kan tilpasses til andre malariaparasitten arter og andre organismer og celletyper.

Figur 1 illustrerer et rutediagram af de vigtigste trin og varigheden af RNA FISH metoden. En række repræsentative billeder af godt og dårligt bevarede farvede mRNA FISH eksperimenter med enkelt P. falciparum IE er vist i figur 2. Denne intracellulære protozo har en lille (fra 1 til 1,5 um diameter) kerne, som er farvet blåt med DAPI. Fireogtyve timer efter erythrocyt invasion i P. falciparum processen med schizogony, dvs nukl...

Watch this Scientific Journal Video about Analysis of Single-cell Gene Transcription by RNA Fluorescent In Situ Hybridization FISH at JoVE.com

Analysis of Single-cell Gene Transcription by RNA Fluorescent In Situ Hybridization FISH

Fluorescent<em&gt; In situ</em&gt; Hybridisering (FISH) til at identificere mRNA transkripter i individuelle celler muliggør analyse af polygen aktivitet såsom den samtidige transskription af mere end ét medlem af<em&gt; Var</em&gt; Multigenfamilie i<em&gt; Plasmodium falciparum</em&gt; Inficerede erythrocytter<sup&gt; 1</sup&gt;. Teknikken er fleksibel og kan anvendes på forskellige typer af gener, celler og organismer.

Analyse af enkeltcelle-gentransskription ved RNA Fluorescent<em&gt; In Situ</em&gt; Hybridisering (FISH)

Adhesion of Plasmodium falciparum infected erythrocytes (IE) to human endothelial receptors during malaria infections is mediated by expression of PfEMP1 ...

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Analysis of Single-cell Gene Transcription by RNA Fluorescent In Situ Hybridization (FISH)

Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization

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Whole mount in situ hybridization is one of the most widely used techniques in developmental biology.  Here, we present a high-resolution double ...

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Knowing the timing, level, cellular localization, and cell type that a gene is expressed in contributes to our understanding of the function of the gene. ...

Detection of Viral RNA by Fluorescence in situ Hybridization (FISH)

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Påvisning af viralt RNA ved fluorescens<em&gt; In situ</em&gt; Hybridisering (FISH)

Viruses  that infect cells elicit specific changes to normal cell functions which serve  to divert energy and resources for viral replication. Many ...

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Kromosom Replikerer Timing Kombineret med Fluorescent<em&gt; In situ</em&gt; Hybridisering

Mammalian DNA replication initiates at multiple sites along chromosomes at different times during S phase, following a temporal replication program. The ...

Whole Mount RNA Fluorescent in situ Hybridization of Drosophila Embryos

Whole Mount RNA Fluorescent in situ Hybridization of Drosophila Embryos

Hele Mount RNA Fluorescent<em&gt; In situ</em&gt; Hybridisering af<em&gt; Drosophila</em&gt; Embryoner

Assessing  the expression pattern of a gene, as well as the subcellular localization  properties of its transcribed RNA, are key features for ...

RNA In situ Hybridization in Whole Mount Embryos and Cell Histology Adapted for Marine Elasmobranchs

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RNA<em&gt; In situ</em&gt; Hybridisering i Hele Mount Embryoner og Cell Histologi Tilpasset til Marine bruskfisk

Marine  elasmobranchs are valued animal models for biomedical and genomic studies as  they are the most primitive vertebrates to have adaptive immunity ...

Visualization and Analysis of mRNA Molecules Using Fluorescence In Situ Hybridization in Saccharomyces cerevisiae

Visualization and Analysis of mRNA Molecules Using Fluorescence In Situ Hybridization in Saccharomyces cerevisiae

Visualisering og analyse af mRNA molekyler under anvendelse Fluorescens<em&gt; In Situ</em&gt; Hybridisering i<em&gt; Saccharomyces cerevisiae</em

The Fluorescence in situ Hybridization (FISH) method allows one to detect nucleic acids in the native cellular environment. Here we provide a protocol for ...

Analysis of Global RNA Synthesis at the Single Cell Level following Hypoxia

Analyse af Global RNA-syntese på enkelt celle niveau efter Hypoxia

Hypoxia or lowering of the oxygen availability is involved in many physiological and pathological processes. At the molecular level, cells initiate a ...