Genejet whole blood genomic dna purification mini kit

Manufactured by Thermo Fisher Scientific

Sourced in United States, Lithuania, Germany

The GeneJET Whole Blood Genomic DNA Purification Mini Kit is a laboratory tool designed for the rapid and efficient extraction of high-quality genomic DNA from whole blood samples. The kit utilizes a simple spin-column procedure to isolate DNA, making it a convenient and reliable solution for downstream genomic analysis applications.

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63 protocols using genejet whole blood genomic dna purification mini kit

Genotyping of Immunomodulatory SNPs

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Peripheral blood was collected from each patient and DNA was extracted using the GeneJET Whole Blood Genomic DNA Purification Minikit (Thermo Scientific, inc) according to the manufacturer’s instructions. Genotyping was done using TaqMan SNP assays for Interleukin-10 SNPs; rs1800872(ID_ C___1747363_10), rs1800896 (ID_ C___1747360_10 ) and TNFa SNP rs1800629(ID_ C___7514879_10) (Applied Biosystems, Foster City, CA).

Eldesouki R.E., Kishk R.M., Abd El-Fadeal N.M., Mahran R.I., Kamel N., Riad E., Nemr N., Kishk S.M, & Mohammed E.A. (2024). Association of IL-10–592 C > A /-1082 A > G and the TNFα -308 G > A with susceptibility to COVID-19 and clinical outcomes. BMC Medical Genomics, 17, 40.

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Assessment of Metabolic Syndrome and PPAR-γ Polymorphisms

Cited 1 time

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Any case or control participant with dermatological diseases other than psoriasis, polycystic ovarian syndrome, coronary artery disease, breast cancer, and/or blood transfusion in the last 6 months was excluded from the study.
Every case and control participant underwent the following steps

Determination of BMI.[34 (link)] Selected cases and controls included obese and nonobese participants

Evaluation for the presence of MS whose criteria were identified according to the National Cholesterol Education Program Adult Treatment Panel III[35 (link)]

Detection of PPAR-γ gene polymorphism by restriction fragment length polymorphism polymerase chain reaction (PCR).

DNA extraction from the whole blood was done using the GeneJET whole blood genomic DNA purification Mini Kit (Thermo Scientific Lithuania). DNA eluted in buffer AE was stored at −20°C for further PCR procedure.

Seleit I., Bakry O.A., Abd El Gayed E, & Ghanem M. (2019). Peroxisome Proliferator-Activated Receptor-γ Gene Polymorphism in Psoriasis and Its Relation to Obesity, Metabolic Syndrome, and Narrowband Ultraviolet B Response: A Case–Control Study in Egyptian Patients. Indian Journal of Dermatology, 64(3), 192-200.

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Multiplex SNP Genotyping by iMLDR

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The genomic DNA was extracted using a GeneJET Whole Blood Genomic DNA Purification Mini Kit (Thermo Scientific, USA) as per the product instruction. The SNP genotyping work was performed using an improved multiplex ligation detection reaction (iMLDR) technique developed by Genesky Biotechnologies Inc. (Shanghai, China). A multiplex PCR-ligase detection reaction method was used in the iMLDR. For each SNP, the alleles were distinguished by different fluorescent labels of allele-specific oligonucleotide probe pairs. Different SNPs were further distinguished by different extended lengths at the 3’end. Two negative controls were set: one with double-distilled water as template and the other with DNA sample without primers while keeping all other conditions the same in one plate. Duplicate tests were designed and the results were consistent. A random sample accounting for ~ 5% (n = 40) of the total DNA samples was directly sequenced using Big Dye-terminator version 3.1 and an ABI3730XL automated sequencer (Applied Biosystems) to confirm the results of iMLDR.

Li Y., Xu X., Zhang D., Cheng W., Zhang Y., Yu B, & Zhang Y. (2019). Genetic variation in the leukotriene pathway is associated with myocardial infarction in the Chinese population. Lipids in Health and Disease, 18, 25.

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Genomic DNA Extraction and Bisulfite Conversion

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GeneJET Whole Blood Genomic DNA Purification Mini Kit (Thermo Fisher Scientific) was used for DNA extraction. Then DNA concentration and purity were determined with the NanoDrop. Bisulfite conversion of the genomic DNA was performed with an EZ DNA Methylation™ Kit following the instruction manual. Finally, 5μl of bisulfite-treated DNA per sample was used to detect methylation using HRM-qPCR.

Zhang B., Zhou T., Wu H., Zhao M, & Lu Q. (2021). Difference of IFI44L methylation and serum IFN-a1 level among patients with discoid and systemic lupus erythematosus and healthy individuals. Journal of Translational Autoimmunity, 4, 100092.

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Whole Blood DNA Extraction

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Genomic DNA was extracted using the Thermo Fisher Scientific GeneJET Whole Blood Genomic DNA Purification Mini Kit (Thermo Fisher Scientific) from 500 μl of whole blood. After extraction, the samples were stored at -20°C until genotyping analysis.

Phoswa W.N., Ramsuran V., Naicker T., Singh R, & Moodley J. (2020). HLA-G Polymorphisms Associated with HIV Infection and Preeclampsia in South Africans of African Ancestry. BioMed Research International, 2020, 1697657.

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Molecular Screening of Plasmodium Parasites in Wild Macaques

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Ninety-three blood samples were collected from wild macaques by Dr. Wirasak Fungfuang in three areas of Thailand from July 2017–July 2019, including samples from wild M. arctoides at the Pa La U waterfall, Huahin district, Prachuap Kiri Khan Province, wild M. leonina at the Khao Yai National Park, Nakornratchasima Province, and wild M. fascicularis at Chang Island, Mu Ko Ranong National Park, Ranong Province. The gDNA was extracted directly from samples using GeneJET Whole Blood Genomic DNA Purification Mini Kit (Thermo Fisher Scientific, USA) and stored at −20°C until use. In our previous study, all gDNA samples showed negative results for dengue virus (DENV), Zika virus (ZIKV), Chikungunya virus (CHIKV), Leptospira spp., and Burkholderia pseudomallei using molecular detection [17 (link),18 (link)]. To analyze Plasmodium species, a nested PCR approach was employed, following the previously described method [19 (link)]. Plasmodium-positive samples underwent further examination using species-specific primers to identify P. knowlesi, P. coatneyi, P. cynomolgi, P. inui, and P. fieldi, as outlined in a previous study [20 (link)] (Table 2) and P. cynomolgi RPA-LFD assay.

Rattaprasert P., Chavananikul C., Fungfuang W., Chavalitshewinkoon-Petmitr P, & Limudomporn P. (2023). Combining isothermal recombinase polymerase amplification with lateral flow assay for diagnosis of P. cynomolgi malaria infection. PLOS Neglected Tropical Diseases, 17(7), e0011470.

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Whole Blood DNA Extraction Kit

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Genomic DNA was extracted following the manufacturer's instructions using a GeneJET Whole Blood Genomic DNA Purification Mini Kit (Thermo Scientific, EU/Lithuania).

Makled A.F., Ali S.A., Eldahdouh S.S., Sleem A.S., Eldahshan M.M., Elsaadawy Y., Salman S.S, & Mohammed Elbrolosy A. (2023). Angiotensin-Converting Enzyme-2 (ACE-2) with Interferon-Induced Transmembrane Protein-3 (IFITM-3) Genetic Variants and Interleukin-6 as Severity and Risk Predictors among COVID-19 Egyptian Population. International Journal of Microbiology, 2023, 6384208.

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VACV DNA Extraction and qPCR Quantification

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VACV DNA was extracted using a GeneJET whole blood genomic DNA purification minikit (Thermo Scientific) according to the manufacturer’s protocol. VACV DNA qPCR was performed using iTaq Universal SYBR green Supermix (Bio-Rad) as described previously (51 (link)). Serial dilutions were included in each qPCR run to develop a standard curve and determine the PCR efficiency of the primer sets in each experiment. qPCR analysis was performed using 10 ng of DNA and 1 μM concentrations of each primer. Viral DNA quantification used primers F-HA and R-HA. The primers are listed in Table 1.

Linville A.C., Rico A.B., Teague H., Binsted L.E., Smith G.L., Albarnaz J.D, & Wiebe M.S. (2022). Dysregulation of Cellular VRK1, BAF, and Innate Immune Signaling by the Vaccinia Virus B12 Pseudokinase. Journal of Virology, 96(11), e00398-22.

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Genomic DNA Purification from Blood

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DNA was extracted from the blood samples using a Gene JET Whole Blood Genomic DNA Purification Mini Kit (Thermofisher Scientific, Waltham, MA, USA) following the guidelines of the manufacturer. Briefly, using a micropipette, a 200 μL blood sample was added to an Eppendorf tube, and then Proteinase K solution (20 μL) was added for protein destruction and release of nucleic acid. Later on, a lysis solution (400 μL) was added, and the product was subjected to vortexing. The sample was then incubated at 56 °C in a water bath for 10 min. Afterward, 200 μL of ethanol was added, and the mixture was shifted to a spin column and subjected to centrifugation at 6000× g for 1 min. The spin column was twice washed with 500 μL of wash buffer and centrifuged (8000× g for 1 min and 12,000× g for 3 min). In the last step, elution buffer (200 μL) was added, and the genomic DNA was collected after centrifugation (8000× g for 1 min). The purified DNA was stored at −20 °C in a microcentrifuge tube until used for further processing.

Atif F.A., Ullah S., Cossío-Bayúgar R., Kashif M., Khan A.U, & Wu W.F. (2023). Molecular Epidemiology, Seasonality and Phylogenetic Investigations of Anaplasma ovis in Small Ruminants from Diverse Agro-Climatic Regions of Punjab, Pakistan. Microorganisms, 11(10), 2430.

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Extraction and Genotyping of DEFB1 SNP

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Extraction of DNA was done using a blood sample through Gene JET™ Whole Blood Genomic DNA Purification Mini Kit (THERMO SCIENTIFIC, EU/Lithuania), following the manufacturer's instructions. SNP for −20G/A (rs11362) DEFB1 gene was performed by PCR-RFLP. Primer’s sequence was: F: CTT GAC TGT GGC ACC TCC CTT CAG-(sense) and R: -CAG CCC TGG GGA TGG GAA ACT C- (antisense). PCR reactions were carried out in a total volume of 30 uL containing 60 ng DNA, 3 µL 10 × PCR Gold Buffer, 2.5 mM MgCl2, 200 uM of each deoxynucleotide triphosphate, 0.4 mM of each primer, and 1 U of Ampli Taq Gold polymerase. Samples were denatured at 95 °C for 10 min followed by 30 cycles of 95 °C for the 60 s, 66 °C hybridization temperature for 60 s and 72 °C for the 60 s, and a final extension for 10 min at 72 °C. After PCR, the products were digested with a specific restriction enzyme, ScrFI (for G-20A) (Jingmei Biotech, Shanghai). Genotyping was performed blindly. The 268-bp PCR product was digested by ScrFI overnight at 37 °C.²¹ (link)

Farag A.G., Shoeib M.A., labeeb A.Z., Sleem A.S., Khallaf H.M., Khalifa A.S., Elshaib M.E., Elnaidany N.F, & Hanout H.M. (2022). Human beta-defensin 1 circulating level and gene polymorphism in non-segmental vitiligo Egyptian patients. Anais Brasileiros de Dermatologia, 98(2), 181-188.

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