Measurement of yield and quality of DNA in human buffy coat is extraction method dependent

Díaz, Tatiana; Ortega-Pinazo, Jesús; Martínez, Beatriz; Jiménez, Álvaro; Gómez‐Zumaquero, J. M.; Caracuel, Z; Hortas, Maria Luisa; Ferro, P.

doi:10.1080/10826068.2022.2119574

Cited by 1 publication

(2 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The QuantiFlour® dsDNA System (Promega, Madison WI, USA) was used to detect and quantify double‐stranded DNA (dsDNA) in plasma samples from patients and healthy volunteers 30 . Thawed plasma samples were centrifuged (20 817 g , 30 min, 4°C) and soluble dsDNA (= cfDNA) was analyzed in the supernatant.…”

Section: Methodsmentioning

confidence: 99%

“…The QuantiFlour® dsDNA System (Promega, Madison WI, USA) was used to detect and quantify double-stranded DNA (dsDNA) in plasma samples from patients and healthy volunteers. 30 Thawed plasma samples were centrifuged (20 817 g, 30 min, 4°C) and soluble dsDNA (= cfDNA) was analyzed in the supernatant. According to manufacturer's instructions, 2 μL of 1:5 pre-diluted plasma was mixed with 200 μL of QuantiFluor® dsDNA Dye working solution and fluorescence (504 nm Ex /531 nm Em ) was measured using the Quantus™ Fluorometer (Promega).…”

Section: Detection Of Cfdna In Plasma Samplesmentioning

confidence: 99%

See 1 more Smart Citation

Clinical relevance of cell‐free DNA during venovenous extracorporeal membrane oxygenation

Lingel,

Haus,

Paschke

et al. 2023

Artificial Organs

View full text Add to dashboard Cite

BackgroundThrombosis remains a critical complication during venovenous extracorporeal membrane oxygenation (VV ECMO). The involvement of neutrophil extracellular traps (NETs) in thrombogenesis has to be discussed. The aim was to verify NETs in the form of cell‐free DNA (cfDNA) in the plasma of patients during ECMO.MethodsA fluorescent DNA‐binding dye (QuantifFluor®, Promega) was used to detect cell‐free DNA in plasma samples. cfDNA concentrations from volunteers (n = 21) and patients (n = 9) were compared and correlated with clinical/technical data before/during support, ECMO end and time of a system exchange.ResultsBefore ECMO, patients with a median (IQR) age of 59 (51/63) years, SOFA score of 11 (10/15), and ECMO run time of 9.0 (7.0/19.5) days presented significantly higher levels of cfDNA compared to volunteers (6.4 (5.8/7.9) ng/μL vs. 5.9 (5.4/6.3) ng/μL; p = 0.044). Within 2 days after ECMO start, cfDNA, inflammatory, and hemolysis parameters remained unchanged, while platelets decreased (p = 0.005). After ECMO removal at the end of therapy, cfDNA, inflammation, and coagulation data (except antithrombin III) remained unchanged. The renewal of a system resulted in known alterations in fibrinogen, d‐dimers, and platelets, while cfDNA remained unchanged.ConclusionDetection of cfDNA in plasma of ECMO patients was not an indicator of acute and circuit‐induced thrombogenesis.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Detection Of Cfdna In Plasma Samplesmentioning

confidence: 99%