Maternal and control plasma was isolated from 10 mL blood samples received in Streck (La Vista, USA) BCT tubes and stored at −80 °C. Cell-free DNA was extracted either from 4 mL of plasma using the QIAsymphony Circulating Nucleic Acid kit (Qiagen, Hilden, Germany) into a final volume of 60 μL AVE buffer or from 2 mL plasma using a QIAamp DSP MiniElute virus kit (Qiagen, Hilden, Germany) into a final volume of 50 μL AVE buffer. Genomic DNA (gDNA) was extracted from leukocytes by QIAsymphony DSP DNA kit (Qiagen, Hilden, Germany).
Qiasymphony circulating nucleic acid kit
Manufactured by Qiagen
Sourced in Germany
The QIAsymphony Circulating Nucleic Acid kit is a product designed for the purification of circulating nucleic acids, such as cell-free DNA and RNA, from various sample types. The kit utilizes magnetic-bead-based technology to facilitate the isolation and purification process. It is intended for use with the QIAsymphony automated sample processing platform.
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Lab products found in correlation
2 protocols using qiasymphony circulating nucleic acid kit
1
Isolation and Extraction of Maternal Cell-Free DNA
2
Quantitative Plasma DNA Purification
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In EDTA tubes, 9-mL blood samples were collected from donors and different cohorts of patients. Plasma was isolated by centrifugation at 2,000 g for 10 minutes within 4 hours and stored at -80°C until use. Plasma was centrifuged again at 10,000 × g for 10 minutes before purification, and cysteine-rich polycomb-like protein 1 (CPP1) DNA fragments were added as exogenous internal control. 13 DNA from patients was purified from 2 × 2 or 2 × 4 mL of plasma on the QIAsymphony SP instrument using the QIAsymphony Circulating Nucleic Acid Kit (Qiagen, Hilden, Germany) according to manufacturer instructions. DNA from healthy donors was purified from 1 × 4 mL of plasma.
DNA was eluted in 60 mL of the supplied buffer and water added to 400 mL. Quantitative PCR was performed on 6 × 3 mL of eluate to quantify CPP1 and total cell-free DNA (gB2M) as previously described. 13 The remaining eluates were concentrated on an Amicon Ultra 0.5 centrifugal filter unit (Millipore, Billerica, MA) to 20 mL.
All participants provided written informed consent.
DNA was eluted in 60 mL of the supplied buffer and water added to 400 mL. Quantitative PCR was performed on 6 × 3 mL of eluate to quantify CPP1 and total cell-free DNA (gB2M) as previously described. 13 The remaining eluates were concentrated on an Amicon Ultra 0.5 centrifugal filter unit (Millipore, Billerica, MA) to 20 mL.
All participants provided written informed consent.
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