Flexigene chemistry

Manufactured by Qiagen

Sourced in Germany

The FlexiGene Chemistry is a nucleic acid purification solution that can be used to extract and purify DNA from a variety of sample types. It provides a flexible and reliable method for DNA isolation.

Automatically generated - may contain errors

Lab products found in correlation

Quantstudio 7 flex detection system, by Thermo Fisher Scientific (2 mentions) 384 well block module, by Thermo Fisher Scientific (2 mentions) Agencourt ampure protocol, by Beckman Coulter (1 mentions) Abi33730xl genetic analyzer, by Thermo Fisher Scientific (1 mentions) Bigdye terminator v3.1 cycle sequencing kit, by Thermo Fisher Scientific (1 mentions) Agencourt cleanseq, by Beckman Coulter (1 mentions) Sequencher 4, by Gene Codes (1 mentions) Taqman snp genotyping assays and chemistries, by Thermo Fisher Scientific (1 mentions) 384well temo liquid handling robots, by Tecan (1 mentions) 7900 ht taqman sequence detector system, by Thermo Fisher Scientific (1 mentions) Genomiphi, by Cytiva (1 mentions) Picogreen fluorescent dye, by Thermo Fisher Scientific (1 mentions)

3 protocols using flexigene chemistry

Genotyping of Alzheimer's Variants

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DNA was extracted from blood using AutoGenFlexStar (AutoGen) and FlexiGene Chemistry (Qiagen) or brain using AutoGen 245 T using standard protocols. Genotyping was performed using TaqMan assays (rs616338, C___2270073_20; rs72824905, C__97909430_10; rs429358, C___3084793_20; rs7412, C____904973_10) following manufacturers protocol, using a QuantStudio 7 Flex Detection System with a 384-Well Block Module (Applied Biosystems, Foster City, CA).

Conway O.J., Carrasquillo M.M., Wang X., Bredenberg J.M., Reddy J.S., Strickland S.L., Younkin C.S., Burgess J.D., Allen M., Lincoln S.J., Nguyen T., Malphrus K.G., Soto A.I., Walton R.L., Boeve B.F., Petersen R.C., Lucas J.A., Ferman T.J., Cheshire W.P., van Gerpen J.A., Uitti R.J., Wszolek Z.K., Ross O.A., Dickson D.W., Graff-Radford N.R, & Ertekin-Taner N. (2018). ABI3 and PLCG2 missense variants as risk factors for neurodegenerative diseases in Caucasians and African Americans. Molecular Neurodegeneration, 13, 53.

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Genotyping of LBD and PSP Patients

Cited 1 time

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All control and a subset of 230 PSP (PSP Series 1) participants had genotypes obtained in our prior study [8 (link)]. We genotyped all 973 LBD-NP and an additional 810 PSP (PSP Series 2) patients using the same methods.
DNA was extracted from blood using AutoGenFlexStar (AutoGen) and FlexiGene Chemistry (Qiagen) or brain using AutoGen 245T using standard protocols. Genotyping was performed using TaqMan assays (ABI3_rs616338, C_2270073_20; PLCG2_rs72824905, C_97909430_10) following manufacturers’ protocol, using a QuantStudio 7 Flex Detection System with a 384-Well Block Module (Applied Biosystems, Foster City, CA).
All minor allele carriers (ABI3_rs616338-T, PLCG2_rs72824905-G) were confirmed using Sanger Sequencing. Polymerase chain reaction (PCR) primers with the following sequences were used to amplify and sequence the genomic region flanking the mutations: ABI3 5′-CTTCCTGCTCGCACCCGAC-3′, 5′-CTAATGCAGCATCCCCAACT-207 3′, PLCG2 5′- CCATAAATGAGGGCTCTCAG-3′, 5′-CATACCCACCTCACCCTTGT-3′. PCR products were purified using the Agencourt AMPure protocol (Beckman Coulter, CA) and sequenced using a Big Dye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems). Sequencing reactions were purified using Agencourt CleanSEQ (Beckman Coulter, CA) and run on an ABI33730xl Genetic Analyzer (Applied Biosystems). Sequences were analyzed using Sequencher 4.8 (Gene Codes Corporation, MI).

Strickland S.L., Morel H., Prusinski C., Allen M., Patel T.A., Carrasquillo M.M., Conway O.J., Lincoln S.J., Reddy J.S., Nguyen T., Malphrus K.G., Soto A.I., Walton R.L., Crook J.E., Murray M.E., Boeve B.F., Petersen R.C., Lucas J.A., Ferman T.J., Uitti R.J., Wszolek Z.K., Ross O.A., Graff-Radford N.R., Dickson D.W, & Ertekin-Taner N. (2020). Association of ABI3 and PLCG2 missense variants with disease risk and neuropathology in Lewy body disease and progressive supranuclear palsy. Acta Neuropathologica Communications, 8, 172.

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Automated DNA Genotyping Platform

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DNA was prepared using the FlexiGene chemistry (Qiagen, Hilden, Germany). For preparation of genotyping, DNA samples were evaluated by gel electrophoresis and adjusted to 20-30 ng/µl DNA content using the Picogreen fluorescent dye (Molecular Probes -Invitrogen, Carlsbad, Ca, USA) on a robotic platform using TECAN liquid handling equipment. One microliter of genomic DNA was amplified with the GenomiPhi (Amersham, Uppsala, Sweden) whole genome amplification kit and fragmented at 99°C for three minutes. Five ng of DNA was performed using TaqMan® SNP Genotyping Assays and chemistries (Applied Biosystems, Foster City, CA, USA) on an automated platform with TECAN Freedom EVO and 384well TEMO liquid handling robots (TECAN, Männedorf, Switzerland) as described before. [12] All process data were logged and administered with a database-driven LIMS. Reactions were completed and read in a 7900 HT TaqMan sequence detector system (Applied Biosystems, Foster City, CA, USA). The amplification reaction was carried out with the TaqMan universal master mix. Thermal cycling conditions consisted of 1 cycle for 10 minutes at 95°C, followed by 45 cycles for 15 seconds at 95°C, and 45 cycles for 1 minute at 60°C.

Stickel F., Buch S., Nischalke H.D., Weiss K.H., Gotthardt D., Fischer J., Rosendahl J., Marot A., Elamly M., Casper M., Lammert F., McQuillin A., Zopf S., Spengler U., Marhenke S., Kirstein M.M., Vogel A., Eyer F., von Felden J., Wege H., Buch T., Schafmayer C., Braun F., Deltenre P., Berg T., Morgan M.Y, & Hampe J. (2018). Genetic variants in PNPLA3 and TM6SF2 predispose to the development of hepatocellular carcinoma in individuals with alcohol-related cirrhosis. The American journal of gastroenterology, 113(10).

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