Dna isolation kit for mammalian blood

Manufactured by Roche

Sourced in United States, Germany

The DNA Isolation Kit for Mammalian Blood is a laboratory product designed to extract and purify DNA from mammalian blood samples. It provides a simple and efficient method for the isolation of high-quality genomic DNA suitable for various downstream applications, such as PCR, sequencing, and genetic analysis.

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Lab products found in correlation

Qubit 3.0 fluorometer, by Thermo Fisher Scientific (6 mentions) Quickgene dna whole blood kit s, by Kurabo (4 mentions) Genomic dna screentape system, by Agilent Technologies (2 mentions) Infinium methylationepic beadchip, by Illumina (2 mentions) Nanodrop 2000, by Thermo Fisher Scientific (2 mentions) Generead dna ffpe kit, by Qiagen (1 mentions) Generead dna quantimize assay kit, by Qiagen (1 mentions) Nucleospin blood l kit, by Macherey-Nagel (1 mentions) Goldengate assay, by Illumina (1 mentions) Tryple, by Thermo Fisher Scientific (1 mentions) Massarray assay design 4, by Labcorp (1 mentions) Massarray platform, by Labcorp (1 mentions) Qiaamp circulating nucleic acid kit, by Qiagen (1 mentions) Nanodrop nd 1000, by Thermo Fisher Scientific (1 mentions) Ampure xp bead, by Beckman Coulter (1 mentions) M220 focused ultrasonicator, by Covaris (1 mentions) Illustrator v 23, by Adobe (1 mentions) Taqman snp genotyping assay, by Thermo Fisher Scientific (1 mentions) Lightcycler 480 2, by Roche (1 mentions) Magna pure lc, by Roche (1 mentions)

23 protocols using dna isolation kit for mammalian blood

Extraction and Purification of Tumor and Leukocyte DNA

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Tumor DNA was extracted and purified by proteinase K digestion and ethanol extraction, followed by a clean-up step on columns (Qiagen) in Cochin cohort and with the GeneRead DNA FFPE Kit (Qiagen) in Wuerzburg cohort, according to the manufacturer’s protocols. The quality of DNA was analyzed by the Nanodrop ND-1000 spectrophotometer (Nyxor Biotech) in Cochin cohort and by the GeneRead DNA QuantiMIZE Assay Kit (Qiagen) in Wuerzburg cohort.
Leukocyte DNA was isolated with the DNA isolation kit for mammalian blood (Roche) in Cochin cohort and with the NucleoSpin Blood L Kit (Macherey-Nagel, Bethlehem, PA, USA) in Wuerzburg cohort, according to the manufacturer’s instructions.

Jouinot A., Lippert J., Fassnacht M., de La Villeon B., Septier A., Neou M., Perlemoine K., Appenzeller S., Sibony M., Gaujoux S., Dousset B., Libe R., Groussin L., Ronchi C.L., Assié G, & Bertherat J. (2020). Intratumor heterogeneity of prognostic DNA-based molecular markers in adrenocortical carcinoma. Endocrine Connections, 9(7), 705-714.

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Genotyping SNPs in T-cell Pathways

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DNA was isolated from peripheral blood leukocytes using the DNA Isolation Kit for Mammalian Blood (Roche, Indianapolis, USA), and stored at 4°C prior to further investigation within 2 days. GoldenGate assay (IlluminaInc, San Diego, USA); capable of multiplexing from 96 to 1,536 SNPs in a single reaction over a 3-day period; was used for SNP genotypingaccording to the manufacturer’s instructions. A 96-SNP GoldenGate assay was designed using SNPs selected from the 26 genes of the T-cell pathways. To ensure the accuracy of the genotyping, quality control was performed using exclusion criteria for SNPs as follows: 1) maximum per-person missing rate>5%; 2) Hardy-Weinberg disequilibrium p-value< 0.001; 3) maximum per-SNP missing rate > 5%; 4) minor allele frequency< 0.01.

Zhang Y., Li J., Wang C, & Zhang L. (2015). Association between the Interaction of Key Genes Involved in Effector T-Cell Pathways and Susceptibility to Developallergic Rhinitis: A Population-Based Case-Control Association Study. PLoS ONE, 10(7), e0131248.

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Quantifying Mitochondrial DNA Copy Number

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Cells were stimulated for 24 h under the same conditions as those used in real-time PCR experiments. After removal of the cell culture medium, the cells were detached using TrypLE (Thermo Fisher Scientific), and clear MEM containing 10% FBS was added. After the cells were collected, they were centrifuged (himac CE15R, Hitachi Koki Co., Ltd., Tokyo, Japan, 15,000× g, 4 °C, 1 min). The supernatant was discarded, and samples were collected for mtDNA copy number measurement.
For DNA extraction, the cells were collected using a DNA isolation kit for mammalian blood (Roche, Basel, Switzerland) according to the manufacturer’s instructions. DNA was extracted from the cells and subjected to real-time PCR. The primer sequences are listed in Table S3. The expression of NADH-ubiquinone oxidoreductase chain 1 (ND1), used as a marker of mtDNA copy number, was normalized to that of glyceraldehyde-3-phosphate dehydrogenase (GAPDH).

Sakurai A., Sakurai T., Ho H.J., Chiba H, & Hui S.P. (2024). Kaempferol Improves Cardiolipin and ATP in Hepatic Cells: A Cellular Model Perspective in the Context of Metabolic Dysfunction-Associated Steatotic Liver Disease. Nutrients, 16(4), 508.

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Tsetse Fly DNA Extraction Protocol

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DNA extraction from all tsetse fly samples analyzed in this study was done by following a protocol adopted for extraction of DNA from crushed tsetse fly samples. Briefly, dried flies in tubes containing stainless beads were transferred to a smashing machine and crushed at 3,000 rpm for 45 sec. DNA from crushed flies was isolated using the DNA Isolation kit for mammalian blood (Roche USA) as per the manufacturer’s protocol with the slight modification suggested for extraction of DNA from Buffy coat, where Red blood cell lysis step is bypassed. This allows lysis of all cells in the solution at once including trypanosomes using the white cell lysis buffer. The DNA sample was stored at -80°C until analysis.

Gaithuma A.K., Yamagishi J., Martinelli A., Hayashida K., Kawai N., Marsela M, & Sugimoto C. (2019). A single test approach for accurate and sensitive detection and taxonomic characterization of Trypanosomes by comprehensive analysis of internal transcribed spacer 1 amplicons. PLoS Neglected Tropical Diseases, 13(2), e0006842.

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Identifying Genetic Factors in Limb Abnormalities

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We identified 253 patients with idiopathic lower limb abnormalities (clubfoot, vertical talus, tibial hemimelia and polydactyly) from the Washington University Musculoskeletal DNA Databank who were recruited from St Louis Children's Hospital and St Louis Shriners Hospital. Patients were excluded from the study if they had additional non-limb congenital anomalies, developmental delay or known underlying etiologies such as arthrogryposis, myelomeningocele or myopathy. DNA was collected from blood and saliva from affected probands and family members after obtaining informed consent. DNA extractions were performed using the manufacturer's protocols using either the DNA Isolation Kit for Mammalian Blood (Roche, Indianapolis, IN, USA) or the Oragene Purifier for saliva (DNA Genotek, Kanata, ON, Canada).

Alvarado D.M., Yang P., Druley T.E., Lovett M, & Gurnett C.A. (2014). Multiplexed direct genomic selection (MDiGS): a pooled BAC capture approach for highly accurate CNV and SNP/INDEL detection. Nucleic Acids Research, 42(10), e82.

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Genotyping of Genomic DNA from Blood Samples

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Blood samples collected from each study subject were stored at -80°C before DNA extraction. Genomic DNA was isolated from peripheral blood leukocytes, using the DNA Isolation Kit for Mammalian Blood (Roche, Indianapolis, USA). Genotyping of the selected SNPs was performed using the Sequenom MassARRAY platform with iPLEX Gold chemistry on a matrix-assisted laser desorption/ ionization time-of-flight mass spectrometer (Sequenom, San Diego, California) according to the supplier's instructions. The extension primers were designed using MassARRAY Assay Design 4.0 software (Sequenom, San Diego, California). Products generated from the polymerase chain reaction (PCR) were transferred to 384-well Spectro-CHIPs (Sequenom, San Diego, California), and analyzed in a Compact Mass Spectrometer, using the MassARRAY Typer 4.0 Software. The PCR assay was arrayed with positive and negative controls and duplicated samples in each 384-well format as quality control. All the genotyping results were generated and checked by laboratory staff blinded to the sample status.

Tan C., Hu W., Huang Y., Zhou J, & Zheng S. (2016). Risk of eighteen genome-wide association study-identified genetic variants for colorectal cancer and colorectal adenoma in Han Chinese. Oncotarget, 7(47), 77651-77663.

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Peripheral Blood Sampling and DNA Isolation

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Peripheral blood (10 mL)was sampled one to three days before the operation. Blood samples in EDTA tubes were centrifuged for 10 minutes at 1600 g at 4°C, and white blood cells were collected and stored. The supernatants from these samples were further centrifuged at 16000 g for 10 minutes at 4°C, and plasma was collected and stored at −80°C until use. White blood cell DNA was isolated using the DNA Isolation Kit for Mammalian Blood (Roche), and cfDNA was isolated using the QIAamp Circulating Nucleic Acid Kit (QIAGEN) according to the manufacturer's protocol. A total of 100–300 ng cfDNA was acquired from 1 mL plasma.

Jiang N., Zhou J., Zhang W., Li P., Liu Y., Shi H., Zhang C., Wang Y., Zhou C., Peng C., Zhang W., Hao Y., Sun Q., Li Y, & Zhao X. (2020). RNF213 gene mutation in circulating tumor DNA detected by targeted next‐generation sequencing in the assisted discrimination of early‐stage lung cancer from pulmonary nodules. Thoracic Cancer, 12(2), 181-193.

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DNA Methylation Profiling Protocol

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Leukocyte DNA was extracted from EDTA blood samples, using the DNA Isolation kit for Mammalian Blood (Roche). DNA quality was assessed on a Genomic DNA ScreenTape system (Agilent) and quantified using a Qubit 3.0 Fluorometer (Thermofisher). DNA was treated by bisulphite and then hybridized to the Infinium MethylationEPIC BeadChip (Illumina; ~850 000 sites), starting from 500 ng of DNA. All experiments were performed following the manufacturer’s instructions at the P3S Post-Genomic Platform of Sorbonne University (Paris, France).

Armignacco R., Jouinot A., Bouys L., Septier A., Lartigue T., Neou M., Gaspar C., Perlemoine K., Braun L., Riester A., Bonnet-Serrano F., Blanchard A., Amar L., Scaroni C., Ceccato F., Rossi G.P., Williams T.A., Larsen C.K., Allassonnière S., Zennaro M.C., Beuschlein F., Reincke M., Bertherat J, & Assié G. (2021). Identification of glucocorticoid-related molecular signature by whole blood methylome analysis. European Journal of Endocrinology, 186(2), 297-308.

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Quantifying DNA-BaP adducts in PBMCs

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Peripheral blood mononuclear cells (PBMCs) were isolated from blood of each volunteer at each time point and DNA isolated
for determination of total [¹⁴C]-BaP_eq bound using the graphite AMS assay as previously described (Felton and Turteltaub, 1994 (link)). DNA (average yield 24 ± 8 μg/mL whole blood) was
extracted with a DNA Isolation Kit for Mammalian Blood (Roche Diagnostics, Indianapolis, IN). The DNA pellet was washed twice with
100% and once with 70% ethanol prior to solubilizing in nuclease-free water for determination of the concentration and purity via
a Nanodrop ND-1000 (Thermo Scientific, Wilmington, DE). Samples were prepared as 200 ng/μL solutions. Twenty μg of
DNA were analyzed by graphite AMS. The limit of adduct detection (LOD) for the 20 μg DNA sample was 0.2 fg (10 fg
[¹⁴C]-BaP adducts/mg DNA) or approximately 0.5 adducts/10¹¹ nucleotides.

Madeen E., Siddens L.K., Uesugi S., McQuistan T., Corley R.A., Smith J., Waters K.M., Tilton S.C., Anderson K.A., Ognibene T., Turteltaub K, & Williams D.E. (2018). Toxicokinetics of Benzo[a]pyrene in Humans: Extensive Metabolism as Determined by UPLC-Accelerator Mass Spectrometry Following Oral Micro-Dosing. Toxicology and applied pharmacology, 364, 97-105.

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Genomic DNA Extraction and Preparation

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Peripheral blood collected in EDTA tubes was first spun down at 4000 × g at 4 degrees Celsius for 10 min. Subsequently, the buffy coat layer was transferred to a 1.5mL Eppendorf and stored at −80 degrees Celsius. Genomic DNA was extracted using the Sigma-Aldrich DNA Isolation Kit for Mammalian Blood (Roche, cat. no. 11667327001) and then sonicated to 150bp using the Covaris M220 Focused-ultrasonicator. Sonicated DNA was then size selected using Agencourt AMPure XP beads (Beckman Coulter, cat. no. A63881) using a 0.8× ratio.

Ul Haq S., Schmid S., Aparnathi M.K., Hueniken K., Zhan L.J., Sacdalan D., Li J.J., Meti N., Patel D., Cheng D., Philip V., Tsao M.S., Cabanero M., de Carvalho D., Liu G., Bratman S.V, & Lok B.H. (2022). Cell-free DNA methylation-defined prognostic subgroups in small-cell lung cancer identified by leukocyte methylation subtraction. iScience, 25(12), 105487.

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