Blood samples were collected from the jugular or cephalic vein from 39 canine patients who presented with acute hemoperitoneum secondary to splenic neoplasm rupture. All dogs underwent preoperative staging, the results of which have been previously reported23 (link). The whole blood was transferred to Cell Free DNA BCT Streck tubes (Streck, Inc.), and processed as per manufacturer’s instructions to isolate plasma. Collected plasma and buffy coat were stored at – 80 °C. Blood sample collection was performed prior to splenectomy and at subsequent clinical visits. Through clinical and histopathological examinations, patients were diagnosed with malignant splenic neoplasms (n = 25) and benign splenic lesions (n = 14). All histopathologic diagnoses were verified through post hoc review of medical records and clinical data to confirm the diagnoses. Blood samples were also collected in Cell Free DNA BCT Streck tubes (Streck, Inc.) from nine healthy dogs and processed similarly.
Cell free dna bct tube
Manufactured by Streck
Sourced in United States, Germany
Cell-Free DNA BCT tubes are a product designed to collect and preserve cell-free DNA samples for analysis. The tubes contain proprietary reagents that stabilize the samples, preventing degradation and inhibiting the release of additional cell-free DNA during storage and transportation.
Automatically generated - may contain errors
Lab products found in correlation
Qiaamp circulating nucleic acid kit, by Qiagen (20 mentions)
Qubit dsdna hs assay kit, by Thermo Fisher Scientific (11 mentions)
Qubit 2.0 fluorometer, by Thermo Fisher Scientific (6 mentions)
Qiasymphony dsp circulating dna kit, by Qiagen (5 mentions)
Qiaamp dna mini kit, by Qiagen (4 mentions)
Maxwell rsc ccfdna plasma kit, by Promega (4 mentions)
Qiasymphony dsp virus pathogen midi kit, by Qiagen (4 mentions)
Magmax cell free dna isolation kit, by Thermo Fisher Scientific (3 mentions)
Qiasymphony sp system, by Qiagen (3 mentions)
Qiamp circulating nucleic acid kit, by Qiagen (3 mentions)
Allprep dna rna mini kit, by Qiagen (3 mentions)
Cellsave tube, by Menarini Silicon Biosystems (2 mentions)
Elution buffer, by Qiagen (2 mentions)
Qiamp blood mini kit, by Qiagen (2 mentions)
Taqman rnase p detection kit, by Thermo Fisher Scientific (2 mentions)
2200 tapestation instrument, by Agilent Technologies (2 mentions)
Ion plus fragment library kit, by Thermo Fisher Scientific (2 mentions)
Edta tube, by BD (2 mentions)
Cell free dna collection tube, by Roche (2 mentions)
Qiasymphony sp, by Qiagen (2 mentions)
121 protocols using cell free dna bct tube
1
Canine Splenic Neoplasm Rupture Evaluation
2
Plasma Isolation from Healthy Elderly
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Blood samples were collected into EDTA tubes (Beckton Dickinson) or Cell-free DNA BCT tubes (Streck). Plasma was separated by centrifugation at 2,500g for 20 minutes at room temperature and stored at -80°C.
Blood specimens from 428 healthy volunteers (214 females and 214 males) were collected into Cell-free DNA BCT tubes (Streck) for a study at the Catholic University of Leuven. Collection of plasma from these asymptomatic donors was approved by the local Institutional Review Board. All volunteers were 65 years or older.
Blood specimens from 428 healthy volunteers (214 females and 214 males) were collected into Cell-free DNA BCT tubes (Streck) for a study at the Catholic University of Leuven. Collection of plasma from these asymptomatic donors was approved by the local Institutional Review Board. All volunteers were 65 years or older.
3
EGFR TKI Therapy Resistance in NSCLC
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Altogether 401 samples of 242 NSCLC patients, previously treated with first- or second-generation EGFR TKI therapy have been analyzed in the Molecular Pathology Laboratory of the National Institute of Oncology between 2019 and 2021. The number of cases in different sample types was as follows: 3 pleural effusions, 5 cell blocks, 6 cytology smears, 6 resection specimens, 17 lung biopsies and 364 blood samples. All the cytology and tissue samples contained viable tumor cells. The average number of blood samples per patient was 1.65; the maximum number was 6 blood samples per patient. Blood specimens and pleural effusions were collected and stored either in K2EDTA S-Monovette tubes (Sarstedt, Nümbrecht, Germany) at 4°C or in Cell-Free DNA BCT Streck tubes (Streck, La Vista, NE, United States) at room temperature, depending on the proximity of the sampling site. In the first case, liquid biopsies were processed within 2 h, while in the case of Streck tubes the time interval did not exceed 72 h.
The study was reviewed and approved by ETT-TUKEB, Health and Scientific Committee of Ministry of Human Resources of Hungary. Number of permission: IV/1792-4/2021/EKU.
The study was reviewed and approved by ETT-TUKEB, Health and Scientific Committee of Ministry of Human Resources of Hungary. Number of permission: IV/1792-4/2021/EKU.
4
Isolation of Circulating Cell-Free DNA
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Circulating cfDNA was extracted from healthy, de‐identified volunteers recruited from the NIEHS Clinical Research Unit (Research Triangle Park). The Institutional Review Board of NIEHS approved the protocol for this study and methodologies conformed to the standards set by the Declaration of Helsinki.
Peripheral whole blood was collected from two healthy male volunteers in Streck Cell‐Free DNA BCT® tubes (La Vista), and the plasma was separated within 2 h of collection by a double centrifugation protocol. The first spin at 1600× g was performed at room temperature for 10 min in a mega‐centrifuge (Sorvall RT7, Thermo Fisher Scientific). Following careful plasma separation, centrifugation was repeated at 16,000× g for 10 min in a benchtop micro‐centrifuge (Eppendorf 5415D, Hauppauge), and the clean plasma was transferred to a cryovial for storage at −80°C until ccfDNA isolation.
Peripheral whole blood was collected from two healthy male volunteers in Streck Cell‐Free DNA BCT® tubes (La Vista), and the plasma was separated within 2 h of collection by a double centrifugation protocol. The first spin at 1600× g was performed at room temperature for 10 min in a mega‐centrifuge (Sorvall RT7, Thermo Fisher Scientific). Following careful plasma separation, centrifugation was repeated at 16,000× g for 10 min in a benchtop micro‐centrifuge (Eppendorf 5415D, Hauppauge), and the clean plasma was transferred to a cryovial for storage at −80°C until ccfDNA isolation.
5
Circulating Tumor Cell and Cell-Free DNA Isolation
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Blood samples were collected in different types of tubes following this order for each patient: CellSave tubes (Menarini Silicon Biosystems, Bologna, Italy) for CMC count, EDTA tubes, and Streck Cell-Free DNA BCT tubes (Streck, La Vista, NE, USA) for cfDNA analysis. Plasma was obtained by double centrifugation (1600 × g for 10 minutes and 16,000 × g for 10 min at room temperature) and stored at −80 °C. The cfDNA was isolated using the QIAamp Circulating Nucleic Acid Kit (Qiagen, Hilden, Germany), and quality control (QC) was performed using 4200 TapeStation system (cfDNA ScreenTape, Agilent Technologies, Santa Clara, CA, USA). QC was passed if the percentage of cfDNA was >82% (region table set from 50 to 700 bp, Supplementary Fig. 1 ). Samples for CMC enumeration were processed within 96 hours from blood collection.
6
Plasma cfDNA Extraction Protocol
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Whole blood was collected in Streck cell-free DNA BCT tubes (La Vista, NE), and stored for no longer than 3 days at room temperature until processing. Collection of plasma was done by spinning the BCT tubes at 1,600 g for 10 min at room temperature, the plasma fraction was then transferred to 2 ml microfuge tubes. The plasma was then spun at 16,000 g for 10 min at room temperature to remove residual cellular debris. The plasma was then carefully transferred to a new 2 ml microfuge tube and stored at −80°C. cfDNA was extracted from 2 ml of plasma using Qiagen QIAamp Circulating Nucleic Acid Kit according to the manufacturer’s instructions, eluted in 50 µl into low bind tubes (1.7 ml Microtube (Maximum Recovery) Cat#22-281LR, Olympus Plastics, Genesee Scientific, El Cajon, CA) and stored at −80°C.
7
Plasma cfDNA Isolation from Whole Blood
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Whole blood was drawn into Streck Cell-Free DNA BCT Tubes (Streck, La Vista, USA, cat. no. 218992). Plasma was separated according to manufacturer’s protocol. Briefly, samples were centrifuged at 1.600 x g for 10 minutes at room temperature followed by a second centrifugation of the supernatant at 16.000 x g for 10 minutes at room temperature. Plasma was stored at −80°C until further processing. cfDNA was isolated from 4 mL plasma using the MagMAX Cell-Free DNA Isolation Kit (Thermo Fisher Scientific, Waltham, USA, cat. no. A29319) either manually or automated on the KingFisher Flex System (Thermo Fisher Scientific, Waltham, USA). The volume after extraction was 36 µL.
8
dd-cfDNA Quantification for Transplant Monitoring
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Venous blood was collected in Streck Cell‐Free DNA BCT tubes before the performance of endomyocardial biopsies and was shipped to the central Clinical Laboratories Improvements Act–certified laboratory at CareDx, Inc. Details of the standardized specimen processing and analytical methods to determine the percentage of dd‐cfDNA (AlloSure®) have been published.10 The targeted next‐generation sequencing assay uses highly polymorphic single nucleotide polymorphisms to quantify dd‐cfDNA without the need for separate genotyping of the recipient or the donor.10 All measurements were performed by laboratory technicians unaware of the clinical identity of the samples.
9
Plasma cfDNA Extraction and Quantification
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Pre- and post-surgical plasma were prospectively collected within a week pre-surgery and 6-weeks post-surgery (Fig 1A ). Pre- and post-surgical plasma was available for all patients but one; CTE029, where only post-surgical blood was available. Platelet-poor plasma was obtained from whole blood drawn into two 10mL STRECK cell-free DNA BCT tubes (STRECK, #218962). The plasma was separated from the buffy coat within 4 hours of collection and centrifuged for 10 minutes at 2k rpm. The plasma was then spun again at 2k rpm for 10 minutes to yield platelet-poor plasma and stored at -80°C until cfDNA extraction. CfDNA was isolated from 1-5mL of platelet-poor plasma using the Circulating Nucleic Acid kit and eluted into 10-30ul of elution buffer (Qiagen, MD, USA; 55114), either following the manufacturer’s protocol or with the following modifications: columns were eluted twice with 150ul elution buffer heated to 37°C. 140ul of each elution was combined and concentrated to 10-30ul using the DNA clean and concentrator-5 kit (Zymo, #DCC-5). CfDNA yield was quantitated using the Qubit dsDNA HS assay kit (Thermo Fisher Scientific, #Q32854) (S4 Table ). Normal control cfDNA (NC cfDNA) from pooled EDTA plasma of cancer-free individuals was processed following the manufacturer’s protocol.
10
Sensitive cfDNA Methylation Assay
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Whole blood was collected in Streck cell-free DNA BCT tubes (La Vista, NE) and stored for no longer than 3 days at room temperature before further processing. The blood plasma collection and storage, the cfDNA extraction and storage, and the cfDNA sodium bisulfite treatment were performed as described earlier [22 (link)]. The set of ten biomarkers that identifies most tumors of common carcinoma types (Additional file 1 : Table S1) and the design of qPCR amplicons specific for the biomarker loci were described before [20 (link), 22 (link)]. The two-step qPCR consisting of 15 cycles of pre-amplification using cocktail of all primer pairs and 50 cycles of loci-specific qPCR detection was done as previously described by our group [22 (link)].
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