Quickgene dna whole blood kit

Manufactured by Kurabo

Sourced in Japan

The QuickGene DNA whole blood kit is a nucleic acid extraction device designed for the purification of genomic DNA from whole blood samples. The kit provides a standardized and efficient method for extracting high-quality DNA for various downstream applications.

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Lab products found in correlation

Taqman snp genotyping assay, by Thermo Fisher Scientific (3 mentions) Fastgene gel pcr extraction kit, by Nippon Genetics (1 mentions) Rtaq dna polymerase, by Takara Bio (1 mentions) Kod fx neo, by Toyobo (1 mentions) 2720 thermal cycler, by Thermo Fisher Scientific (1 mentions) Genelute mammalian genomic dna miniprep kit, by Merck Group (1 mentions) Qiamp dna blood midi kit, by Qiagen (1 mentions) Allelic discrimination taqman snp genotyping system, by Thermo Fisher Scientific (1 mentions) Peak scanner software, by Thermo Fisher Scientific (1 mentions)

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QuickGene whole blood kit (2 citations)

7 protocols using quickgene dna whole blood kit

Mutant A3B DNA Amplification and Cloning

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HEK293T cells were transfected with pDON-EGFP, pEF-UGI, and expression vectors for A3B WT or its mutants, using the XtremeGENE HP DNA Transfection Reagent. After two-day cultures, total DNA was extracted using the Quick Gene DNA whole blood kit (KURABO). First round PCR was performed using the primers and rTaq DNA polymerase (Takara), with the following reaction profile: 30 s at 94 °C, 25 cycles of 30 s at 94 °C, 40 s at 62 °C, and 90 s at 72 °C, followed by 10 min at 72 °C^{20 (link)}. The amplicons were separated by electrophoresis in 1% (w/v) agarose gel, and extracted with the FastGene Gel/PCR Extraction kit (NIPPON Genetics). We used 100 ng of first-round PCR products as templates for nested PCR using KOD Fx Neo (TOYOBO), with the following reaction profile: 5 min at 95 °C, 30 cycles of 10 s at 81–88 °C, 30 s at 62 °C, and 60 s at 72 °C, followed by 5 min at 72 °C. The amplicons derived at each of the indicated denaturation temperatures were then cloned into the pT7-blue vector (Novagen).

Matsumoto T., Shirakawa K., Yokoyama M., Fukuda H., Sarca A.D., Koyabu S., Yamazaki H., Kazuma Y., Matsui H., Maruyama W., Nagata K., Tanabe F., Kobayashi M., Shindo K., Morishita R., Sato H, & Takaori-Kondo A. (2019). Protein kinase A inhibits tumor mutator APOBEC3B through phosphorylation. Scientific Reports, 9, 8307.

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Genotyping of rs53576 Polymorphism

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Genomic DNA was extracted from blood, drawn after the T0 measurement, using the Quick Gene DNA whole blood Kit (Kurabo, Japan). Genotyping of the rs53576 was performed using the TaqMan SNP Genotyping assay (Applied Biosystems; C___3290335_10). The TaqMan genotyping reaction was amplified on an Applied Biosystem 2720 Thermal Cycler 95 °C for 15 min, and 95 °C for 15 sec, and 60 °C for 1 min, for 40 cycles) and fluorescence was detected by using the Applied Biosystem 7500 Real-Time PCR systemon (Thermo Fisher Scientific Inc.). To assess genotyping reproducibility, a random ~5% selection of the sample were re-genotyped for all SNPs; all genotypes matched initially designated genotypes.

Bornemann B., Kovacs P, & Singer T. (2019). Voluntary upregulation of heart rate variability through biofeedback is improved by mental contemplative training. Scientific Reports, 9, 7860.

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Genotyping of Genetic Variants

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Genomic DNA was extracted from blood using the Quick Gene DNA whole blood Kit (Kurabo, Japan). Genotyping of the two previously reported SNPs rs8102595 (A/G) and rs3826795 (G/A)11 (link) was performed using the TaqMan SNP Genotyping assay (Applied Biosystems; C _29247492_10; C_31640839_10). To assess genotyping reproducibility, a random ~5% selection of the sample were re-genotyped for all SNPs; all genotypes matched initial designated genotypes. Potential functional significance of the studied genetic variants was checked using the Regulome Database, which includes public datasets from GEO, the ENCODE project, and published literature20 (link).

Pfeiffer S., Krüger J., Maierhofer A., Böttcher Y., Klöting N., El Hajj N., Schleinitz D., Schön M.R., Dietrich A., Fasshauer M., Lohmann T., Dreßler M., Stumvoll M., Haaf T., Blüher M, & Kovacs P. (2016). Hypoxia-inducible factor 3A gene expression and methylation in adipose tissue is related to adipose tissue dysfunction. Scientific Reports, 6, 27969.

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Bone Marrow Transplantation in JAK2V617F Mice

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Recipient female C57BL/6 J mice aged between 8 and 10 weeks (Charles River Japan, Inc.) were lethally irradiated with a total dose of 9.0 Gy 24 h before BMT^{22 (link)}. Whole BM cells were harvested from donor femurs and tibiae. The cells were washed with PBS and 5.0 × 10⁶ of BM cells were injected in the recipient mice via the tail vein. Peripheral blood parameters and chimerism were analyzed at 4 weeks after transplantation and at the termination of the experiments. DNA was isolated using the QuickGene DNA whole blood kit (KURABO) and quantitative PCR was performed using THUNDERBIRD SYBR qPCR Mix with the following primers; forward primer for donor and recipients, 5ʹ-CTTTCTTCGAAGCAGCAAGCATGA-3ʹ, reverse primer for recipients; 5ʹ-CTGGCTTTACTTACTCTCCTCTCCACAGAC-3ʹ reverse primer for donors; 5ʹ-AACCAGAATGTTCTCCTCTCCACAGAA-3ʹ. Delta Ct (Ct_donor– Ct_total) was calculated to estimate Jak2V617F VAF in JAK2^V617F-BMT mice.

Kimishima Y., Misaka T., Yokokawa T., Wada K., Ueda K., Sugimoto K., Minakawa K., Nakazato K., Ishida T., Oshima M., Koide S., Shide K., Shimoda K., Iwama A., Ikeda K, & Takeishi Y. (2021). Clonal hematopoiesis with JAK2V617F promotes pulmonary hypertension with ALK1 upregulation in lung neutrophils. Nature Communications, 12, 6177.

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Genotyping of CRTC1 SNPs in Sorbs

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Genotype data of 13 CRTC1 SNP markers were extracted from a genome wide SNP data set available for 887 non-diabetic individuals from the German Sorbs population [22 (link)]. The SNP variant rs7256986 was de novo genotyped in all 3 study cohorts. Genomic DNA was extracted from whole blood tissue samples in the Sorbs (N = 887) and replication cohort (N = 314) using the QIAmp DNA Blood Midi Kit (Qiagen Inc., Valencia, CA, USA) or QuickGene DNA Whole Blood Kit (Kurabo, Japan). Genomic DNA was extracted from adipose tissue in the Adiposity cohort (N = 168) using the GenEluteTM Mammalian Genomic DNA Miniprep Kit (SIGMA-ALDRICH, USA) according to the manufacturer's protocols. SNP data were obtained by using Allelic Discrimination TaqMan SNP Genotyping System (Applied Biosystems by Life-Technologies Carlsbad, CA, USA). Fluorescence signals were detected by the ABI 7500 Real-Time PCR system. Genotyping errors were excluded by random re-genotyping (5% of all samples) while all genotypes matched with initially obtained results. Further we used water as non-template controls (N = 6 per run). All SNPs were in Hardy–Weinberg equilibrium (all P < 0.05).

Rohde K., Keller M., la Cour Poulsen L., Rønningen T., Stumvoll M., Tönjes A., Kovacs P., Horstmann A., Villringer A., Blüher M, & Böttcher Y. (2019). (Epi)genetic regulation of CRTC1 in human eating behaviour and fat distribution. EBioMedicine, 44, 476-488.

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Microsatellite Genotyping Protocol

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DNA was extracted using QuickGene DNA whole blood kit (KURABO). Eight microsatellite loci, D10S190, D10S191, D18S21, BAT40, BAT25, BAT26, D2S123, and D5S346, were amplified by PCR from genomic DNA. For each microsatellite locus, the forward primer was fluorescently labeled with FAM, VIC or NED (Applied Biosystems, Foster City, CA). PCR reaction and analysis by capillary electrophoresis using ABI 3100 xl Genetic Analyzer (Applied Biosystems) were performed by SystemBiotics Inc. (Kanagawa, Japan). Data were processed using the Peak Scanner Software (Applied Biosystems). The primer sequences for PCR amplification of each microsatellite marker were: D10S190 (forward 5′-FAM-GTGTTTGGGTCATGGAGATG-3’/reverse 5′-AGGCAAAGCAGGAGCA-3′), D10S191 (5′-VIC-CTTTAATTGCCCTGTCTTC-3’/5′-TTAATTCGACCACTTCCC-3′), D18S21 (5′-VIC-GTGGTTATTGCCTTGAAAAG-3’/5′-GATGACATTTTCCCTCTAG-3′), BAT40 (5′-NED-ACAACCCTGCTTTTGTTCCT-3’/5′-GTAGAGCAAGACCACCTTG-3′), BAT25 (5′-VIC-TCGCCTCCAAGAATGTAAGT-3’/5′-TCTGGATTTTAACTATGGCTC-3′), BAT26 (5′-FAM-TGACTACTTTTGACTTCAGCC-3’/5′-GTTTCTAACCATTCAACATTTTTATCCC-3′), D2S123 (5′-NED-AAACAGGATGCCTGCCTTTA-3’/5′-GGACTTTCCACCTATGGGAC-3′), D5S346 (50-FAM-ACTCACTCTAGTGATAAATCGGG-3’/5′-AGCAGATAAGACAGTATTACTAGTT-3′).

Roberts B.T., Ying C.Y., Gautier J, & Maller J.L. (1999). DNA replication in vertebrates requires a homolog of the Cdc7 protein kinase. Proceedings of the National Academy of Sciences of the United States of America, 96(6), 2800-2804.

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Genotyping of BMI-Associated SNP rs8050136

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Genomic DNA was extracted from blood samples using the QuickGene DNA whole blood kit (Kurabo, Japan). Genotyping of the SNPs rs8050136 was performed using the TaqMan SNP Genotyping assay (Applied Biosystems; C_2031259_10). To assess genotyping reproducibility, a random ~5 % selection of the sample were re-genotyped for all SNPs; all genotypes matched initial designated genotypes. Rs8050136 belongs to one L.D. group, which includes the previously reported BMI-associated SNP rs9939609 2 .

Krüger J., Berger C., Weidle K., Schleinitz D., Tönjes A., Stumvoll M., Blüher M., Kovacs P, & Klöting N. (2019). Metabolic effects of genetic variation in the human REPIN1 gene. International journal of obesity (2005), 43(4).

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