Nucleospin8 blood isolation kit

Manufactured by Macherey-Nagel

Sourced in Germany

The NucleoSpin8 Blood Isolation kit is a product designed for the rapid and efficient extraction of DNA from whole blood samples. It utilizes a silica-based membrane technology to capture and purify DNA, providing a reliable and consistent method for obtaining high-quality genetic material.

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Lab products found in correlation

Qubit 2.0 fluorometer, by Thermo Fisher Scientific (4 mentions) Nanodrop, by Thermo Fisher Scientific (4 mentions) Qubit dsdna br assay kit, by Thermo Fisher Scientific (2 mentions) Nanodrop spectrophotometer, by Thermo Fisher Scientific (2 mentions) Puregene blood core kit, by Qiagen (1 mentions) Qiaamp dna blood maxi kit, by Qiagen (1 mentions)

6 protocols using nucleospin8 blood isolation kit

Genetic Profiling of Diabetic and Idiopathic Neuropathy

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In total, 166 samples from patients diagnosed with diabetic and idiopathic neuropathy were tested by MIPs-NGS and TSCA-NGS. For 70 of these patients, exons and exon-flanking intron sequences of SCN9A, SCN10A, and SCN11A genes were analyzed by Sanger sequencing to validate MIPs-NGS and TSCA-NGS methods [23 (link)]. Local Medical Ethical Committees of Fondazione IRCCS Istituto Neurologico "Carlo Besta" (Italy), Maastricht University Medical Center (the Netherlands), University of Manchester (United Kingdom) and the Deutsche Diabetes Forschungsgesellschaft EV (Germany) approved this study. Informed consent for genetic testing was given by patients to participate in this study.
Genomic DNA was extracted from peripheral blood by using QIAamp DNA Blood Maxi Kit, Puregene^® Blood Core Kit (Qiagen, Hilden, Germany) or NucleoSpin^®8 Blood Isolation kit (Macherey-Nagel, Düren, Germany). Quality and concentration of the DNA was determined by NanoDrop (Thermo Scientific, Wilmington, USA), and Qubit^® 2.0 Fluorometer using the Qubit^® dsDNA BR assay kit (Life technologies, Bleiswijk, The Netherlands). Isolated DNA was stored with a unique numeric code in the central DNA bank at Maastricht University Medical Centre and IRCCS Foundation “Carlo Besta” Neurological Institute.

Almomani R., Marchi M., Sopacua M., Lindsey P., Salvi E., de Koning B., Santoro S., Magri S., Smeets H.J., Martinelli Boneschi F., Malik R.R., Ziegler D., Hoeijmakers J.G., Bönhof G., Dib-Hajj S., Waxman S.G., Merkies I.S., Lauria G., Faber C.G, & Gerrits M.M. (2020). Evaluation of molecular inversion probe versus TruSeq® custom methods for targeted next-generation sequencing. PLoS ONE, 15(9), e0238467.

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Blood DNA Isolation Protocol

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Genomic DNA was from blood samples using a NucleoSpin8 Blood Isolation kit (Macherey-Nagel, Düren, Germany) or a QIAamp DNA Blood Maxi Kit, Puregene^® Blood Core Kit (Qiagen, Hilden, Germany). DNA extraction was performed according to the manufacturers’ instructions and stored in −20 °C.

Ślęczkowska M., Almomani R., Marchi M., Salvi E., de Greef B.T., Sopacua M., Hoeijmakers J.G., Lindsey P., Waxman S.G., Lauria G., Faber C.G., Smeets H.J, & Gerrits M.M. (2022). Peripheral Ion Channel Genes Screening in Painful Small Fiber Neuropathy. International Journal of Molecular Sciences, 23(22), 14095.

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DNA Extraction from Peripheral Blood

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A peripheral blood sample was taken from all patients, and genomic DNA was extracted from peripheral blood by using QIAamp DNA Blood Maxi Kit/Puregene® Blood Core Kit (Qiagen, Hilden, Germany) or NucleoSpin ®8 Blood Isolation kit (Macherey-Nagel, Düren, Germany). Quality and concentration of the DNA were determined by NanoDrop (Thermo Scientific, Wilmington, DE, USA) and Qubit® 2.0 Fluorometer using the Qubit® dsDNA BR assay kit (Life technologies, Bleiswijk, The Netherlands). Isolated DNA was stored with a unique numeric code in the central DNA bank at Maastricht University Medical Centre and IRCCS Foundation “Carlo Besta” Neurological Institute.

Alsaloum M., Estacion M., Almomani R., Gerrits M.M., Bönhof G.J., Ziegler D., Malik R., Ferdousi M., Lauria G., Merkies I.S., Faber C.G., Dib-Hajj S, & Waxman S.G. (2019). A gain-of-function sodium channel β2-subunit mutation in painful diabetic neuropathy. Molecular Pain, 15, 1744806919849802.

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DNA Extraction from Blood and Saliva

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Genomic DNA was extracted from whole blood or saliva samples using a NucleoSpin8 Blood Isolation kit (Macherey-Nagel, Düren, Germany) or a QIAamp DNA Blood Maxi Kit, Puregene ® Blood Core Kit l (Qiagen, Hilden, Germany). DNA isolation was performed according to the manufacturer's instructions and stored at -20 • C. Quality and quantity checks of extracted DNA were performed using agarose gel, a Qubit 2.0 Fluorometer (Thermo Fisher, San Jone, CA, USA), and a Nanodrop Spectrophotometer (Thermo Fisher, San Jone, CA, USA).

Misra K., Ślęczkowska M., Santoro S., Gerrits M.M., Mascia E., Marchi M., Salvi E., Smeets H.J.M., Hoeijmakers J.G.J., Martinelli Boneschi F.G., Filippi M., Lauria Pinter G., Faber C.G, & Esposito F. (2024). Broadening the Genetic Spectrum of Painful Small-Fiber Neuropathy through Whole-Exome Study in Early-Onset Cases. International journal of molecular sciences, 25(13).

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Genomic DNA Extraction from Whole Blood

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Genomic DNA was extracted from whole blood using the NucleoSpin8 Blood Isolation Kit (Macherey-Nagel, Düren, Germany) or QIAamp DNA Blood Maxi Kit, Puregene^® Blood Core Kit (Qiagen, Hilden, Germany), according to the manufacturer’s instructions at Maastricht UMC+, and IRCCS Neurological Institute Carlo Besta. DNA samples were coded and stored in the central DNA bank at Maastricht UMC+ and IRCCS Neurological Institute Carlo Besta in −20 °C.

Ślęczkowska M., Almomani R., Marchi M., de Greef B.T., Sopacua M., Hoeijmakers J.G., Lindsey P., Salvi E., Bönhof G.J., Ziegler D., Malik R.A., Waxman S.G., Lauria G., Faber C.G., Smeets H.J, & Gerrits M.M. (2022). Peripheral Ion Channel Gene Screening in Painful- and Painless-Diabetic Neuropathy. International Journal of Molecular Sciences, 23(13), 7190.

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Genetic Screening of Voltage-gated Sodium Channels

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Genomic DNA was extracted from whole blood using Nucle-oSpin8 Blood Isolation kit (Macherey-Nagel, Düren, Germany) according to manufacturer's instructions. Coding exons and exon-flanking intronic regions of SCN9A, SCN10A and SCN11A were amplified by PCR and sequenced by Sanger sequencing. Sequences were compared with reference sequence GRCh37. Variants detected were annotated according to guidelines of the Human Genome Variation Society (http://www. hgvs. org/ mutnomen/). Variants which were located in functional domain of the protein and/or at a highly conserved amino acid in mammalian paralogues/human VGSC orthologues were classified and reported according the Practice Guidelines of the Association for Clinical Genetic Science (ACGS) and recommendations of Waxman et al. 22 23 Cosegregation of (potentially) pathogenic variants with the disease was tested, if possible, in cases with a positive family history.

Eijkenboom I., Sopacua M., Hoeijmakers J.G.J., de Greef B.T.A., Lindsey P., Almomani R., Marchi M., Vanoevelen J., Smeets H.J.M., Waxman S.G., Lauria G., Merkies I.S.J., Faber C.G, & Gerrits M.M. (2019). Yield of peripheral sodium channels gene screening in pure small fibre neuropathy. Journal of neurology, neurosurgery, and psychiatry, 90(3).

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