Name: Dijital Damla PCR (ddPCR) kullanılarak toprakta Mikrobiyal Biyokütlenin Belirlenmesi (Video) | JoVE Journal
Uploaded: 2026-04-24T14:20:00.000+00:00
Duration: 7 min 46 s
Description: Öğrenin, toprakta bakteri ve mantar biyokütlesini ddPCR ile mutlak niceliksel olarak belirlemeyi. Bu yöntem, 16S ve 18S rRNA genlerini hedefleyerek mikrobiyal karbon ve hücre sayısı ölçümü sağlar.

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<li>Carini, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Relic+DNA+is+abundant+in+soil+and+obscures+estimates+of+soil+microbial+diversity%5BTitle%5D)+AND+%22Nat+Microbiol%22%5BJournal%5D)">Relic DNA is abundant in soil and obscures estimates of soil microbial diversity.</a> Nat Microbiol. 2 (3), 16242(2016).</li>
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</ol>

<table><tbody><tr><td>Bettersizer S3 Plus&nbsp;</td><td>Bettersize, Çin</td><td>BT-803</td><td></td></tr><tr><td>Santrifüj</td><td>Termoscientif&nbsp;</td><td>SL16&nbsp;</td><td></td></tr><tr><td>Kloroform&nbsp;</td><td>TermoBilimsel</td><td>L14759. AU&nbsp;</td><td></td></tr><tr><td>ddPCR 96-Kuyulu Plakalar</td><td>Bio-Rad, Watford, Birleşik Krallık</td><td>12001925</td><td></td></tr><tr><td>ddPCR Damla Okuyucu Yağı&nbsp;</td><td>Bio-Rad, Watford, Birleşik Krallık</td><td>1863004</td><td></td></tr><tr><td>Dessicator&nbsp;</td><td>Davisil</td><td></td><td></td></tr><tr><td>DG8 Karuş Tutucu</td><td>Bio-Rad, Watford, Birleşik Krallık</td><td>1863051</td><td></td></tr><tr><td>QX200/QX100 Damla Jeneratörü için DG8 Kartuşları</td><td>Bio-Rad, Watford, Birleşik Krallık</td><td>1864008</td><td></td></tr><tr><td>QX200/QX100 Damla Jeneratörü için DG8 Contaları</td><td>Bio-Rad, Watford, Birleşik Krallık</td><td>Q33265</td><td></td></tr><tr><td>DNeasy Power Soil pro kiti &nbsp;</td><td>Qiagen, Almanya</td><td>47017</td><td></td></tr><tr><td>çift zincirli (dsDNA) Geniş Aralıklı (BR) test kiti</td><td>Invitrogen, Almanya</td><td>Q33230</td><td></td></tr><tr><td>FastPrep-24 5G&nbsp;</td><td>MP Biyomedikal Şirketi, Birleşik Krallık</td><td>116005500</td><td></td></tr><tr><td>GridION&nbsp;</td><td>Oxford Nanopore Teknolojileri, Birleşik Krallık</td><td>GRD-MK1CAPX</td><td></td></tr><tr><td>K2SO4</td><td>Merck, Birleşik Krallık</td><td>7778-80-5</td><td></td></tr><tr><td>Mag-Bind Manyetik Boncuklar</td><td>Omega Bio-tek, ABD</td><td>M1378-01</td><td></td></tr><tr><td>MinION akış hücresi</td><td>Oxford Nanopore Teknolojileri, Birleşik Krallık</td><td>FLO-MIN114</td><td></td></tr><tr><td>Boğuk fırın</td><td>Karbolit&nbsp;</td><td>AAF1100</td><td></td></tr><tr><td>Yerel barkod kiti 24</td><td>Oxford Nanopore Teknolojileri, Birleşik Krallık</td><td>SQK-NBD114.24</td><td></td></tr><tr><td>PCR Plaka Isı Sızdırmazlığı, folyo, delinebilir ve s.nbsp;</td><td>Bio-Rad, Watford, Birleşik Krallık</td><td>1814040</td><td></td></tr><tr><td>PX1 PCR Plaka Kapatıcı</td><td>Bio-Rad, Watford, Birleşik Krallık</td><td>1814000</td><td></td></tr><tr><td>Kubit florometre</td><td>Invitrogen, Almanya</td><td>Q33226</td><td></td></tr><tr><td>QX200 Damlacıklı Dijital PCR Sistemi</td><td>Bio-Rad, Watford, Birleşik Krallık</td><td>1864001</td><td></td></tr><tr><td>QX200 ddPCR EvaGreen Supermix</td><td>Bio-Rad, Watford, Birleşik Krallık</td><td>186-4033</td><td></td></tr><tr><td>EvaGreen&nbsp için QX200 Damla Üretim Yağı;</td><td>Bio-Rad, Watford, Birleşik Krallık</td><td>1864005</td><td></td></tr><tr><td>QXDx Damla Jeneratörü&nbsp;</td><td>Bio-Rad, Watford, Birleşik Krallık</td><td>12001049</td><td></td></tr><tr><td>SENSOQUEST LABCYCLER&nbsp;</td><td>Geneflow, Almanya</td><td>1120280125</td><td></td></tr></tbody></table>

dijital damla pcr (ddpcr) kullanılarak toprakta mikrobiyal biyokütlenin belirlenmesi

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

Dijital Damla PCR (ddPCR) kullanılarak toprakta Mikrobiyal Biyokütlenin Belirlenmesi

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

Research

JoVE Journal

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Çevre

Basitleştirilmiş (analog) bir okuma ile nükleik asitleri ölçmek için bir uç nokta dijital testi tanımlıyoruz. Damlacık bazlı dijital tahlillerin toplu floresansını özel enstrümantasyon yerine standart bir qPCR makinesi kullanarak ölçüyoruz ve sonuçlarımızı mikroskopi ile doğruluyoruz.

Dijital Nükleik Asit Kantifikasyon Tahliller Basit Toplu Saati

Bu yazıda, aynı anda Enterococcus spp ölçmek için kullanılabilecek bir çift yönlü dijital PCR deneyi. Ve eğlence sularda genel olarak insan ilişkili dışkı kirlenme göstergeleri olarak HF183 genetik işaretleyiciler açıklanmaktadır.

Eşzamanlı Niceleme için Dubleks Dijital PCR Testi<em&gt; Enterococcus spp.</em&gt; Ve Waters İnsan Fekal ilişkili HF183 Marker

Boya bazlı bir kimya ve damlacık dijital PCR teknolojisi kullanılarak hücresiz mikroRNA'ların miktar tayini için hassas ve doğru bir yöntem açıklanmaktadır.

MicroRNA Niceleme DNA bağlayıcı Boya Kimya ve Damlacık Dijital PCR kullanarak Sirkülasyon

Bitki patojenleri yerinde, özellikle toprak kaynaklı patojenler, hızlı ve doğru tespiti daha fazla inoculum üretim ve alanında bitki hastalıklarının yayılması önlemek için çok önemli olduğunu. Burada bir taşınabilir gerçek zamanlı PCR algılama sistemi kullanılarak geliştirilen yöntem yerinde tanı alan koşullarında sağlar.

Toprak kaynaklı Phytopathogens bir taşınabilir gerçek zamanlı PCR sistemi kullanarak yerinde moleküler tespiti

Burada, toprak, rizosferde ve kök endosphere microbiomes amplicon sıra verileri elde etmek için bir protokol açıklayın. Bu bilgiler kompozisyon ve çeşitlilik bitki ilişkili mikrobiyal toplulukların araştırmak için kullanılan ve bitki türlerinin geniş bir kullanım için uygundur.

Kök Microbiome keşfetmek: toprak, rizosferde ve kök Endosphere bakteriyel topluluk verileri ayıklama

Burada açıklanan serbestçe kullanılabilir araçları kullanarak 16S rRNA metagenomik sıralama ve analiz kullanarak mikrobiyom profilleme için basitleştirilmiş bir standart işletim prosedürüdür. Bu protokol mikrobiyom alanında yeni araştırmacılar yanı sıra daha yüksek bir çözünürlükte bakteriyel profilleme elde etmek için yöntemler güncellemeleri gerektiren yardımcı olacaktır.

İki adımlı PCR ve Yeni Nesil 16S rRNA Gen Sıralaması Kullanarak Mikrobiyota Analizi

Burada, tek hücrelerde mutlak mitokondriyal (mt) DNA kopya sayısını ve mtDNA delesyon heteroplazmi seviyelerini ölçmek için bir protokol sunuyoruz.

Dijital Damlacık Polimeraz Zincir Reaksiyonu Kullanarak Tek Hücreli Mitokondriyal DNA Kopya Sayısını ve Heteroplazmiyi Ölçme

Burada, gen tedavisi vektörlerinin klinik gelişimini desteklemek için damlacık dijital polimeraz zincir reaksiyonu ile insan gözyaşlarında adeno ilişkili viral vektörlerin uyumlu tespitinde iyi laboratuvar uygulamalarının geliştirilmesi ve doğrulanması için bir protokol sunuyoruz.

Gözyaşlarının Biyodökülme Çalışmalarında Adeno-İlişkili Viral Vektörlerin Tespiti için Doğrulanabilir Bir Damlacık Dijital Polimeraz Zincir Reaksiyonu Testi

Adeno-ilişkili virüs (AAV) vektör genom kopyalarının kesin miktar tayini kritiktir, ancak standart bir protokol henüz oluşturulmamıştır. Bu protokol, saflaştırılmış AAV numunelerinin hazırlanması ve viral genom titresinin güvenilir bir şekilde ölçülmesi için dijital damlacık polimeraz zincir reaksiyonunun (dd_PCR) yürütülmesi için doğrulanmış bir yöntemi tanımlar.

Dijital Damlacık Polimeraz Zincir Reaksiyonu ile Saflaştırılmış Vektör Örneklerinde Adeno İlişkili Viral Genomların Miktar Tayini

Sahada kullanılabilir DNA analizinin güvenilir ve doğru formları birçok sektör için hayati öneme sahiptir. Bu makale, alanda çevresel mikrobiyal analiz için iki yöntem sunmaktadır: 1) biyolojik ekstraksiyon saha kiti kullanılarak çevresel örneklerden mikrobiyal DNA'nın izolasyonu ve 2) alan qPCR ünitesi aracılığıyla DNA analizi.

Biyolojik Çıkarma Saha Kiti ve Saha QPCR Birimi kullanılarak Alanda Mikrobiyal DNA Analizi

Öğrenin, toprakta bakteri ve mantar biyokütlesini ddPCR ile mutlak niceliksel olarak belirlemeyi. Bu yöntem, 16S ve 18S rRNA genlerini hedefleyerek mikrobiyal karbon ve hücre sayısı ölçümü sağlar.

Dijital Damla PCR (ddPCR) kullanılarak toprakta Mikrobiyal Biyokütlenin Belirlenmesi

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