Innuprep blood dna mini kit

Manufactured by Analytik Jena

Sourced in Germany

The InnuPREP Blood DNA Mini Kit is a laboratory equipment designed for the rapid and efficient extraction of genomic DNA from whole blood samples. The kit utilizes a silica-based membrane technology to purify DNA, which can then be used for various downstream applications such as PCR, sequencing, or other analysis methods.

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Lab products found in correlation

Big dye terminator kit v3, by Thermo Fisher Scientific (1 mentions) Rotor gene q, by Qiagen (1 mentions) Methyl primer express software v1, by Thermo Fisher Scientific (1 mentions) Taqman snp genotyping assay, by Thermo Fisher Scientific (1 mentions) Agilent 8453, by Agilent Technologies (1 mentions) Euro ea 3000, by Eurovector (1 mentions) Elx800 bio tek, by Agilent Technologies (1 mentions) Silica gel 60 f254, by Merck Group (1 mentions) Ion reporter software, by Thermo Fisher Scientific (1 mentions) 3500 series genetic analyzer, by Thermo Fisher Scientific (1 mentions) Bsawi, by New England Biolabs (1 mentions) Multigene, by Labnet (1 mentions)

Spelling variants of this product

InnuPREP DNA Mini Kit (26 citations) InnuPREP Blood DNA Mini kit IPC16 (2 citations)

10 protocols using innuprep blood dna mini kit

Genotyping of AF-associated SNVs

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For all participants, DNA was extracted from peripheral blood samples using the commercially available innuPREP Blood DNA Mini Kit (Analytik Jena AG, Jena, Germany) [9 (link)].
Eight SNVs previously reported to be associated with AF in genome-wide association studies (GWAS) and with a minor allele frequency (MAF) of >5% in Europeans (gnomAD v.2.0.1) were selected for genotyping (Table 1). The selected SNVs were not in strong linkage disequilibrium.
Genotyping was carried out by real-time PCR and high-resolution melting analysis using Rotor-Gene Q (QIAGEN N.V., Venlo, The Netherlands), as described previously [9 (link)]. Primers are available upon request. To confirm the genotyping results, 8–16 samples with different genotypes were randomly selected for Sanger sequencing using the BigDye Terminator Kit v3.1 (Thermo Fisher Scientific, Waltham, MA, USA).

Vogel S., Rudaka I., Rots D., Isakova J., Kalējs O., Vīksne K, & Gailīte L. (2021). A Higher Polygenic Risk Score Is Associated with a Higher Recurrence Rate of Atrial Fibrillation in Direct Current Cardioversion-Treated Patients. Medicina, 57(11), 1263.

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DNA Extraction and Genotyping Protocol

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Genomic DNA was obtained from 250 μL of EDTA-anticoagulated venous blood using innuPREP Blood DNA Mini Kit (Analytik Jena AG, Jena, Germany), according to the manufacturer’s recommendations. The SNP and VNTR genotyping were carried out using locus-specific PCR as described previously [49 (link),50 (link),51 (link),52 (link),53 (link),54 (link)].

Rafikova E., Shadrina M., Slominsky P., Guekht A., Ryskov A., Shibalev D, & Vasilyev V. (2021). SLC6A3 (DAT1) as a Novel Candidate Biomarker Gene for Suicidal Behavior. Genes, 12(6), 861.

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Rapid Genomic DNA Extraction from Blood

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Genomic DNA was extracted from whole blood using the innuPREP Blood DNA Mini Kit from Analytik Jena, Upland, CA, USA) (Catalogue No: AJG#845-KS-1020050). This kit allows the rapid isolation of genomic DNA from EDTA blood samples. The extraction procedure is based on the lysis of the blood sample followed by the binding of the nucleic acids onto the spin filter membrane. After several washing steps, the nucleic acids are eluted from the membrane using elution buffer. The samples were checked for purity and concentration after the extraction procedure. Samples with good purity and concentration were stored under −20 °C until PCR was performed.

Mohanraj J., D’Souza U.J., Fong S.Y., Karkada I.R, & Jaiprakash H. (2022). Association between Leptin (G2548A) and Leptin Receptor (Q223R) Polymorphisms with Plasma Leptin, BMI, Stress, Sleep and Eating Patterns among the Multiethnic Young Malaysian Adult Population from a Healthcare University. International Journal of Environmental Research and Public Health, 19(14), 8862.

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Bisulfite Sequencing of Il36a Promoter

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Genomic DNA was isolated using innuPREP Blood DNA Mini kit (Analytik Jena, Jena, Germany) according to the manufacturer’s instructions. The methylation profile of the proximal Il36a promoter was performed by bisulphite sequencing as described previously with minor modifications [49 (link)]. In brief, 4 μg of genomic DNA was digested with BglII, purified by phenol-chloroform extraction and 500-1000 ng digested DNA was bisulphite-converted as described. Primers were designed using Methyl Primer Express^® Software v1.0 (Applied Biosystems/ThermoFisher Scientific) to amplify specific regions of the genome following bisulphite conversion. The Il36a promoter region from −439 to −112 relative to the transcriptional start site on the (-)-strand was amplified using the primers Il36a_BSP_for (5′-GGAGGGTTTGTTAAGTATTTGT-3′) and Il36a_BSP_rev (5′-AATATCCACTAAAATCAACCTAAAA-3′). For bisulphite sequencing, PCR products were gel-purified and cloned into the pCR2.1-Topo Vector System (ThermoFisher Scientific) and sequenced using the M13rev primer (5′-CAGGAAACAGCTATGAC-3′). Sequencing results were analyzed using QUMA software [50 (link)]. Samples with conversion rate < 90% and sequences identity < 70% were excluded from the analysis. The minimum number of clones for each sequenced condition was ≥10.

Nerlich A., Janze N, & Goethe R. (2021). Analysis of Il36a induction by C/EBPβ via a half-CRE•C/EBP element in murine macrophages in dependence of its CpG methylation level. Genes and Immunity, 22(7-8), 313-321.

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Camel Blood Sampling and DNA Extraction

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Blood samples were collected from the jugular vein of 100 camels from Abu Simbel near the Egypt–Sudan border. All camels were adult males imported from Sudan and kept in quarantine during the sampling process. The blood samples were collected on Na-EDTA tubes and DNA was extracted using innuPREP Blood DNA Mini Kit (Analytik Jena AG, Jena, Germany) following the manufacturer’s recommendations and stored at −20 °C until analyzed.

Mohamed W.M., Ali A.O., Mahmoud H.Y., Omar M.A., Chatanga E., Salim B., Naguib D., Anders J.L., Nonaka N., Moustafa M.A, & Nakao R. (2021). Exploring Prokaryotic and Eukaryotic Microbiomes Helps in Detecting Tick-Borne Infectious Agents in the Blood of Camels. Pathogens, 10(3), 351.

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SNP Genotyping of Blood DNA

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Genomic DNA was obtained from 250 μL of EDTA- anticoagulated venous blood using innuPREP Blood DNA Mini Kit (Analytik Jena AG, Germany), according to the manufacturer's recommendations. For SNP genotyping, we used a set of reagents from TaqMan® SNP Genotyping Assay (Applied Biosystems, USA): C__11592758_10 (rs6264), C___2184734_10 (rs5443), C___1202883_20 (rs1801133), C___3084793_20 (rs429358), and C____904973_10 (rs7412). The In/Del ACE gene polymorphism was determined as described previously 7 (link), 8 (link).

Bondarenko E., Shadrina M., Grishkina M., Druzhkova T., Akzhigitov R., Gulyaeva N., Guekht A, & Slominsky P. (2016). Genetic Analysis of BDNF, GNB3, MTHFR, ACE and APOE Variants in Major and Recurrent Depressive Disorders in Russia. International Journal of Medical Sciences, 13(12), 977-983.

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DNA-Compound Interaction Spectroscopy

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The interaction with DNA of the test compound was studied by using the UV spectrophotometer method reported in literature with small modifications [32 (link)]. In this assay, an innuPREP Blood DNA Mini Kit (Analytik Jena) (AJ Innuscreen GmbH, Berlin, Germany) was used for the extraction of genomic DNA from human blood. The assay was conducted on a PDA spectrophotometer (Agilent 8453) Agilent Technologies, Inc., UK. Methanol and water, in a ratio of 9:1, were used as a blank and solvent to dissolve the compounds. The spectrum of the pure compound was recorded, and, to this, DNA was added in different concentrations, step wise, starting from lowest concentration to higher concentrations, keeping the compound concentration in the reaction mixture constant. The absorbance measurements were recorded by a UV-visible spectrophotometer by keeping the concentrations of compounds constant (100 µM) in the sample cell while varying the concentrations of DNA (0.5 × 10⁻⁶ to 1 × 10⁻⁶, 0.5 × 10⁻⁵ and 1 × 10⁻⁵ M) in the sample cell. The spectra were recorded in the form of spectral peaks in each step and the change in the absorbance was noted. To achieve an equilibrium between the compound and the DNA, the solutions were allowed to stay for at least 5 min before each measurement was made.

Rafique B., Kalsoom S., Sajini A.A., Ismail H, & Iqbal M. (2022). Synthesis, Characterization, Biological Evaluation and DNA Interaction Studies of 4-Aminophenol Derivatives: Theoretical and Experimental Approach. Molecules, 27(4), 1352.

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Spectroscopic Characterization of Schiff Bases

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Melting points were determined in open capillaries on a FALC Instrument SRL360D. TLC (Silica Gel 60 F254) from Merck (Beijing, China) was used to check the compounds’ purities. Elemental analyses were carried out by using Euro EA 3000 (EUROVECTOR, Pavia, Italy) instrument. The infrared spectra of the Schiff bases were recorded on FT-IR (Bruker Optics GmbH & Co. Ettlingen, Germany) with an ATR model (ALPHA 200488) in the frequency range of 4000–550 cm^–1. NMR spectra were recorded in DMSO-d₆ and CDCl₃ solvent on a BRUKER AV-500 MHz NMR (Bruker Beijing Scientific Technology, Beijing, China). A high-resolution multinuclear FT-NMR spectrometer was used in the presence of SiMe₄ (internal standard) at δ = 0 ppm at 25 °C.
The sbsorbance of α-amylase and α-glucosidase activities (at 540 and 405 nm, respectively) were measured on a microplate reader (BioTek Elx-800, Agilent Technologies, Santa Clara, CA, USA). An InnuPREP Blood DNA Mini Kit (Analytik Jena) (AJ Innuscreen GmbH, Berlin, Germany) was used for the extraction of genomic DNA from human blood. A PDA spectrophotometer (Agilent 8453) Agilent Technologies, Berkshire, UK, along with a quartz optical cell (with an effective absorption path of 1 cm), was used for DNA interaction studies of the synthesized Schiff bases.

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Genetic Analysis of Epidermolysis Bullosa

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Genomic DNA was isolated from the peripheral blood of the patient and his parents using an innuPrep Blood DNA mini kit (Analytik Jena, Jena, Deutschland). EB panel sequencing including the COL7A1 gene and at least 16 genes involved in classical Epidermolysis bullosa [28 (link)] was performed on 10ng DNA of the patient with a Personal Genome Machine (PGM) of the Next Generation Sequencing Platform of Life Technologies according to the protocol of the company and analysed with the Ion Reporter Software (Life Technologies, Carlsbad, CA, USA). The mutation in exon 3 and the variant in exon 84 were amplified using PCR (Promega, Madison, GA, USA), confirmed via Sanger sequencing using a 3500 Series Genetic Analyzer (Applied Biosystems, Waltham, MA, USA), and compared with the reference sequence of COL7A1 (NM_000094) using the BLAST algorithm.

Klausegger A., Jeschko N., Grammer M., Cemper-Kiesslich J., Neuhuber F., Diem A., Breitenbach-Koller H., Sander G., Kotzot D., Bauer J.W, & Laimer M. (2022). Recessive Dystrophic Epidermolysis bullosa due to Hemizygous 40 kb Deletion of COL7A1 and the Proximate PFKFB4 Gene Focusing on the Mutation c.425A>G Mimicking Homozygous Status. Diagnostics, 12(10), 2460.

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Genetic Polymorphism Detection in Blood

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Genomic DNA was isolated from blood samples, taken in ethylenediaminetetraacetic acid, using an innuPREP Blood DNA Mini Kit (Analytik Jena AG, Thuringia, Germany). Determination of the SOD and CAT polymorphisms was achieved by restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) analysis using the primers and the restriction enzymes shown in Table 1. PCR was carried out in a final volume of 25 µL, using 1 µL 100 ng genomic DNA, 0.2 U Taq DNA polymerase (Analytik Jena AG), 2.5 µL 10X buffer, 0.2 mM dNTP, 0.4 µM of each of the primers, and 1.5 mM MgCl 2 . PCR cycles were carried out in a DNA Thermal Cycler (MultiGene, Labnet) according to the following program: an initial denaturation of DNA at 94°C for 5 min; 35 cycles of 1 min at 94°C; 30 s at 59°C, or 57°C, or 60°C to detect Cu/Zn-SOD +35 A/C, Mn-SOD T47C, or CAT -21 A/T polymorphisms, respectively; and 45 s at 72°C. A final extension was performed at 72°C for 7 min. The resulting PCR products were digested with the corresponding restriction endonucleases: 5 U HhaI (Thermo Scientific), 2.5 U BsawI (New England BioLabs), and 5 U HinfI (Jena Bioscience) according to the manufacturer recommendations. Digestion products were analyzed electrophoretically on 3% agarose gel stained with ethidium bromide.

Abaidi H., Denden S., Ghazouani A., Trimèche A., Snoussi C., Haj Khelil A., Ben Chibani J, & Hamdaoui M.H. (2015). Mn-SOD 47 CC genotype in combination with high tea consumption may prevent complications in Tunisian type-2 diabetes. Genetics and molecular research : GMR, 14(3).

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