The blood samples were collected in 10 ml Streck Cell-free DNA BCT blood collection tubes (Streck, La Vista, NE, USA) and centrifuged within two hours of blood draw. The literature describes Streck Cell-free DNA BCT blood collection tubes (Streck, La Vista, NE, USA) as suitable for ccfDNA isolation due to the preservative they contain, which prevents leukocyte lysis17 (link). The blood samples were processed within 24 h of blood draw and samples with visible leukozyte lysis (red color of plasma) were excluded from further processing. Blood samples were centrifuged at 1600 × g for 10 min to separate lymphocytes from plasma. Subsequently, the supernatant was centrifuged for another 10 min at 16,000 × g to clear the plasma of remaining debris. The supernatant was stored in 1 ml fractions at 80 °C until use.
Cell free dna bct blood collection tubes
Manufactured by Streck
Sourced in United States
Cell-Free DNA BCT blood collection tubes are designed to stabilize cell-free DNA in whole blood samples. The tubes contain proprietary reagents that preserve the integrity of cell-free DNA during storage and transportation.
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Lab products found in correlation
7 protocols using cell free dna bct blood collection tubes
1
ccfDNA Isolation from Blood Samples
2
CSF Sampling for Molecular Testing
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Between December 2016 and May 2018, we collected CSF samples from 27 patients with cancer who underwent lumbar puncture as part of their routine clinical management. All patients signed informed consent for the analysis of their clinical data within a broad prospective registry conducted at Clinical Cancer Center of the Ludwig Maximilian University Munich (“The informative patient”), which was approved by the local ethics committee according to the Declaration of Helsinki. All patients were discussed in an interdisciplinary tumor board before and after molecular testing, had a clinical indication for molecular testing, and were informed about the purpose of the molecular analysis by the treating physician. CSF samples were collected in 10-ml Cell-Free DNA BCT blood collection tubes (Streck, La Vista, NE, USA) to prevent the release of genomic DNA from cells within the sample and the absorption of cell-free (CF) nucleic acids (NA).
3
Liquid Biopsy Profiling of NSCLC
4
Liquid Biopsy Profiling of NSCLC
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ctDNA analysis was completed using a commercially available targeted next-generation sequencing (NGS) assay (Guardant360; Guardant Health, Redwood City, CA) that is CLIA certified, College of American Pathologists accredited, and New York State Department of Health approved. ctDNA isolation and analytic methods have been described previously.21 (link),22 (link) Briefly, two 10-mL tubes of whole blood were collected in Cell-Free DNA BCT blood collection tubes (Streck, Inc., La Vista, NE) prior to treatment, every two cycles (every 8 weeks), and at progression. At the time of this study, Guardant360 included targeted analysis of single-nucleotide variants in 68 to 70 clinically relevant cancer genes, as well as small insertions/deletions, gene fusions, and copy number gains (CNGs) in a subset of genes including EGFR and common resistance mechanisms to EGFR TKIs (including EGFR CNG, MET CNG, EGFR T790M, oncogenic fusions, BRAF, and HER2 CNG). The variant allele fraction (VAF) was calculated by dividing the number of tumor-derived DNA molecules at each location by the total number of unique ctDNA molecules at the given nucleotide position. We used 5% VAF-detectable EGFRex20ins ctDNA at baseline to analyze clinical outcomes based on a previous association of 5% ctDNA or above with overall survival in NSCLC.23 (link)
5
Plasma Isolation from Whole Blood
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Plasma was isolated within five days from blood draw. An average of 9.5 mL whole blood was collected in Cell-free DNA BCT® blood collection tubes (Streck) via a double centrifugation protocol consisting of a first centrifugation step at 1342 × g for 30 minutes, transfer of the plasma fraction to a secondary tube and a second centrifugation step at 2267 × g for 20 minutes. Plasma was stored at −80 C until further processing.
6
Maternal Plasma cfDNA Sequencing
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Five milliliters of maternal peripheral blood were collected into Streck Cell Free DNA BCT® blood collection tubes (Streck, La Vista, NE, United States). Two hundred microliters of maternal plasma were used for cfDNA extraction, library construction and sequencing as described previously (29 (link)). Briefly, end repairing and adding a non-template dA tail to cfDNA were carried out and DNA amplification products were obtained by polymerase chain reaction (PCR). Then, the amplified double-stranded DNA was thermally denatured to single-strand DNA, cyclized and made into DNA nanoballs (DNBs). Finally, the DNBs were loaded onto chips and sequenced on the BGISEQ-500 sequencing platform (BGI, Shenzhen, China). The average depth of coverage is 0.1X.
7
Retrospective Analysis of Maternal Plasma DNA
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This was a retrospective study. A total of 24,702 participants were retrospectively analyzed from the Women and Children’s Hospital from January 2013 to April 2019 in Xiamen of Fujian Province, China. All the participants provided written informed consent. This study followed strict confidentiality regulations on privacy protection and was approved by the Institutional Review Board of the Women and Children’s Hospital (KY-2019-037). Five milliliters of peripheral blood were taken from the pregnant woman and stored in Streck Cell Free DNA BCT ® blood collection tubes (Streck, La Vista, Nebraska, United States), and plasma DNA was extracted within four days after collection.
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