Name: Bepaling van microbiële biomassa in de bodem met behulp van Digital Droplet PCR (ddPCR) (Video) | JoVE Journal
Uploaded: 2026-04-24T14:20:00.000+00:00
Duration: 7 min 46 s
Description: Leer een geoptimaliseerde ddPCR-workflow voor het kwantificeren van bacteriële en fungale biomassa in bodemmonsters. Deze methode maakt absolute kwantificering van 16S en 18S rRNA-genen mogelijk voor nauwkeurige schatting van microbiele biomassa.

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</ol>

<table><tbody><tr><td>Bettersizer S3 Plus&nbsp;</td><td>Bettersize, China</td><td>BT-803</td><td></td></tr><tr><td>Centrifuge</td><td>Thermoscientific&nbsp;</td><td>SL16&nbsp;</td><td></td></tr><tr><td>Chloroform&nbsp;</td><td>ThermoScientific</td><td>L14759.AU&nbsp;</td><td></td></tr><tr><td>ddPCR 96-Well Plates</td><td>Bio-Rad, Watford, UK</td><td>12001925</td><td></td></tr><tr><td>ddPCR Droplet Reader Oil&nbsp;</td><td>Bio-Rad, Watford, UK</td><td>1863004</td><td></td></tr><tr><td>Dessicator&nbsp;</td><td>Davisil</td><td></td><td></td></tr><tr><td>DG8 Cartridge Holder</td><td>Bio-Rad, Watford, UK</td><td>1863051</td><td></td></tr><tr><td>DG8 Cartridges for QX200/QX100 Droplet Generator</td><td>Bio-Rad, Watford, UK</td><td>1864008</td><td></td></tr><tr><td>DG8 Gaskets for QX200/QX100 Droplet Generator</td><td>Bio-Rad, Watford, UK</td><td>Q33265</td><td></td></tr><tr><td>DNeasy Power Soil pro kit&nbsp;</td><td>Qiagen, Germany</td><td>47017</td><td></td></tr><tr><td>double-stranded (dsDNA) Broad range (BR) assay kit</td><td>Invitrogen, Germany</td><td>Q33230</td><td></td></tr><tr><td>FastPrep-24 5G&nbsp;</td><td>MP Biomedicals, UK</td><td>116005500</td><td></td></tr><tr><td>GridION&nbsp;</td><td>Oxford Nanopore Technologies, UK</td><td>GRD-MK1CAPX</td><td></td></tr><tr><td>K2SO4</td><td>Merck, UK</td><td>7778-80-5</td><td></td></tr><tr><td>Mag-Bind Magnetic beads</td><td>Omega Bio-tek, USA</td><td>M1378-01</td><td></td></tr><tr><td>MinION flow cell</td><td>Oxford Nanopore Technologies, UK</td><td>FLO-MIN114</td><td></td></tr><tr><td>Muffled furnace</td><td>Carbolite&nbsp;</td><td>AAF1100</td><td></td></tr><tr><td>Native barcoding kit 24</td><td>Oxford Nanopore Technologies, UK</td><td>SQK-NBD114.24</td><td></td></tr><tr><td>PCR Plate Heat Seal, foil, pierceable&nbsp;</td><td>Bio-Rad, Watford, UK</td><td>1814040</td><td></td></tr><tr><td>PX1 PCR Plate Sealer</td><td>Bio-Rad, Watford, UK</td><td>1814000</td><td></td></tr><tr><td>Qubit Fluorometer</td><td>Invitrogen, Germany</td><td>Q33226</td><td></td></tr><tr><td>QX200 Droplet Digital PCR System</td><td>Bio-Rad, Watford, UK</td><td>1864001</td><td></td></tr><tr><td>QX200 ddPCR EvaGreen Supermix</td><td>Bio-Rad, Watford, UK</td><td>186-4033</td><td></td></tr><tr><td>QX200 Droplet Generation Oil for EvaGreen&nbsp;</td><td>Bio-Rad, Watford, UK</td><td>1864005</td><td></td></tr><tr><td>QXDx Droplet Generator&nbsp;</td><td>Bio-Rad, Watford, UK</td><td>12001049</td><td></td></tr><tr><td>SENSOQUEST LABCYCLER&nbsp;</td><td>Geneflow, Germany</td><td>1120280125</td><td></td></tr></tbody></table>

bepaling van microbiële biomassa in de bodem met behulp van digital droplet pcr (ddpcr)

Determination of Microbial Biomass in Soil using Digital Droplet PCR (ddPCR)

Bepaling van microbiële biomassa in de bodem met behulp van Digital Droplet PCR (ddPCR)

Soil represents the largest terrestrial pool of organic carbon, yet more than 133 Gt of carbon has been lost from soils over the past two centuries. ...

determination-microbial-biomass-soil-using-digital-droplet-pcr

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Er wordt een gevoelige en nauwkeurige methode beschreven voor de kwantificering van celvrije microRNA's met behulp van een kleurstof-gebaseerde chemie en druppel digitale PCR-technologie.

Circulerende MicroRNA Kwantificering Met behulp van DNA-bindende Dye Chemistry and Droplet Digital PCR

Snelle en nauwkeurige detectie van plant ziekteverwekkers on-site, met name bodem overgedragen ziekteverwekkers, is van cruciaal belang om te voorkomen dat verdere entmateriaal productie en verspreiding van plantenziekten in het veld. De methode ontwikkeld die hier met behulp van een draagbaar real-time PCR detectiesysteem kunt on-site diagnose onder veldomstandigheden.

On-Site moleculaire detectie van bodem-gedragen planteziekteverwekkers met behulp van een draagbare Real-Time PCR-systeem

Hier beschrijven we een protocol om amplicon reeks gegevens van bodem, rhizosfeer en wortel endosphere microbiomes te verkrijgen. Deze informatie kan worden gebruikt voor het onderzoeken van de samenstelling en de diversiteit van de plant-geassocieerde microbiële gemeenschappen, en is geschikt voor het gebruik met een breed scala van plantensoorten.

Verkennen van de wortel Microbiome: extraheren van gegevens van de bacteriële Gemeenschap uit de bodem, de rhizosfeer, en de wortel Endosphere

Hier beschreven is een vereenvoudigde standaard werkings procedure voor microbiome profilering met behulp van 16S rRNA metagenomic sequencing en analyse met behulp van vrij beschikbare instrumenten. Dit protocol zal onderzoekers die nieuw zijn in het microbiome veld en degenen die updates vereisen over methoden om bacteriële profilering bij een hogere resolutie te bereiken helpen.

Microbiota-analyse met behulp van twee-stap PCR en de volgende generatie 16S rRNA Gensequencing

Hier presenteren we een protocol voor het meten van absolute mitochondriale (mt) DNA-kopienummer en mtDNA-deletie heteroplasmieniveaus in enkele cellen.

Het meten van eencellig mitochondriaal DNA-kopienummer en heteroplasmie met behulp van digitale druppelpolymerasekettingreactie

Hier presenteren we een protocol voor de ontwikkeling en validatie van goede laboratoriumpraktijken in de conforme detectie van adeno-geassocieerde virale vectoren in menselijke tranen door druppel digitale polymerasekettingreactie ter ondersteuning van de klinische ontwikkeling van gentherapievectoren.

Een valideerbare druppel digitale polymerasekettingreactietest voor de detectie van adeno-geassocieerde virale vectoren in biosheddingstudies van tranen

Nauwkeurige kwantificering van kopieën van het vectorgenoom van adeno-geassocieerd virus (AAV) is van cruciaal belang, maar er moet nog een gestandaardiseerd protocol worden opgesteld. Dit protocol beschrijft een gevalideerde methode voor het bereiden van gezuiverde AAV-monsters en het uitvoeren van een digitale druppelpolymerasekettingreactie (dd_PCR) om de virale genoomtiter betrouwbaar te kwantificeren.

Kwantificering van adeno-geassocieerde virale genomen in gezuiverde vectormonsters door digitale druppelpolymerasekettingreactie

Betrouwbare en nauwkeurige vormen van veldinzetbare DNA-analyse zijn cruciaal voor veel industrieën. Dit artikel biedt twee methoden voor milieumicrobiële analyse in het veld: 1) de isolatie van microbieel DNA uit omgevingsmonsters met behulp van een biologische extractie-veldkit, en 2) de analyse van DNA via de field qPCR-eenheid.

Microbieel DNA-onderzoek in het veld met behulp van een biologische extractie-veldkit en een veld-qPCR-eenheid

Leer een geoptimaliseerde ddPCR-workflow voor het kwantificeren van bacteriële en fungale biomassa in bodemmonsters. Deze methode maakt absolute kwantificering van 16S en 18S rRNA-genen mogelijk voor nauwkeurige schatting van microbiele biomassa.

Bepaling van microbiële biomassa in de bodem met behulp van Digital Droplet PCR (ddPCR)

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