Ez 96 dna methylation gold kit

Manufactured by Zymo Research

Sourced in United States, Germany

The EZ-96 DNA Methylation-Gold Kit is a product designed for the bisulfite conversion and purification of DNA samples. It is a high-throughput solution for DNA methylation analysis.

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Lab products found in correlation

Infinium humanmethylation450 beadchip, by Illumina (6 mentions) Infinium humanmethylation450 beadchip microarrays, by Illumina (4 mentions) Qiaamp dna mini kit, by Qiagen (4 mentions) Genomestudio software, by Illumina (4 mentions) Nanodrop 1000 spectrophotometer, by Thermo Fisher Scientific (3 mentions) Nanodrop, by Thermo Fisher Scientific (3 mentions) Wizard genomic dna purification kit, by Promega (3 mentions) Humanmethylation450 beadchip, by Illumina (3 mentions) Gotaq hot start green master mix, by Promega (3 mentions) Methylation 450k kit, by Illumina (3 mentions) Allprep dna rna ffpe kit, by Qiagen (3 mentions) Dneasy kit, by Qiagen (3 mentions) Infinium methylationepic beadchip, by Illumina (2 mentions) Hiscan, by Illumina (2 mentions) Microlab star automated liquid handling workstation, by Hamilton Company (2 mentions) Ficoll paque plus, by GE Healthcare (2 mentions) Mini amp dna kit, by Qiagen (2 mentions) Iscan reader, by Illumina (2 mentions) Freedom evo robot, by Tecan (2 mentions) Spectramaxgeminixs spectrophotometer, by Thermo Fisher Scientific (2 mentions)

Spelling variants of this product

EZ-96 DNA Methylation Kit (246 citations) Methylation-Gold kit (45 citations) EZ DNA Methylation-Gold (29 citations) EZ-96 Methylation Kit (6 citations) EZ-96 DNA Methylation-Gold MagPrep Kit (5 citations) Zymo EZ-96 DNA Methylation-Gold Kit™ (2 citations)

95 protocols using ez 96 dna methylation gold kit

Blood Temperature Influences DNA Methylation

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According to our unpublished data, the temperature of blood at processing influences the levels of DNA methylation. Thus, peripheral whole blood was collected by ethylenediaminetetraacetic acid (EDTA) tubes and kept at 4°C within 8 h before being stored at −80°C until further usage. Genomic DNA was isolated from peripheral whole blood using the Genomic DNA Extraction Kit (Rebece, Nanjing, China). DNA was bisulfite converted by EZ-96 DNA Methylation Gold Kit according to the standard protocol (Zymo Research, Orange County, United States).

Yin Q., Yang X., Li L., Xu T., Zhou W., Gu W., Ma F, & Yang R. (2020). The Association Between Breast Cancer and Blood-Based Methylation of S100P and HYAL2 in the Chinese Population. Frontiers in Genetics, 11, 977.

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SOCS-1 DNA Methylation Analysis

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DNA bisulfite conversion was first performed for SOCS-1 gene methylation analysis using the EZ-96 DNA Methylation-Gold Kit (Zymo Research) protocol.

Tuncel F.C., Serin I., Pehlivan S., Oyaci Y, & Pehlivan M. (2022). Epigenetic and genetic investigation of SOCS-1 gene in patients with multiple myeloma. Blood Research, 57(4), 250-255.

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Genome-wide DNA methylation profiling

Cited 1 time

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We extracted the genomic DNA (500 ng) from the saliva of the whole sample, however only 47 subjects paired by age, sex and different anxiety disorder trajectories due to logistical and financial limitations were included in the methylation analysis. The samples were treated with sodium bisulphite using the EZ-96 DNA Methylation-Gold Kit (Zymo Research, Orange, CA, USA) according to the manufacturer’s protocol. DNA methylation status was performed using the Infinium HumanMethylation450 (IHM450) BeadChip^{72 (link)} which covers 99% of Ref Seq genes regions and 96% of CpG islands/CpG island regions to explore the genome-wide DNA methylome^{72 (link)–74 (link)}. Data regarding pre-processing of raw data of IHM450 BeadChip and differential methylation analysis can be found elsewhere^{39 (link)}. All data of DNA methylation including methylated vs. unmethylated probes are deposited in GEO (GSE78975).

Cerveira de Baumont A., Hoffmann M.S., Bortoluzzi A., Fries G.R., Lavandoski P., Grun L.K., Guimarães L.S., Guma F.T., Salum G.A., Barbé-Tuana F.M, & Manfro G.G. (2021). Telomere length and epigenetic age acceleration in adolescents with anxiety disorders. Scientific Reports, 11, 7716.

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Bisulfite Conversion and Amplification

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DNA extracted from each sample was bisulfite converted by EZ-96 DNA Methylation Gold Kit (Zymo Research, Orange, USA) according to the manufacturer’s instructions, and further amplified by bisulfite-specific primers (

Table S1

). After bisulfite treatment, all non-methylated cytosine (C) bases in CpG sites were converted to uracil (U), whereas all methylated C bases remained C. There are no single nucleotide polymorphisms (SNPs) or CpG sites in the primers and in the measured CpGs.

Qiao R., Di F., Wang J., Wei Y., Zhang Y., Xu T., Wang Y., Gu W., Han B, & Yang R. (2020). The Association Between RAPSN Methylation in Peripheral Blood and Early Stage Lung Cancer Detected in Case–Control Cohort. Cancer Management and Research, 12, 11063-11075.

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Whole Blood DNA Extraction and Methylation

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All the peripheral blood samples were collected by EDTA blood collection tubes and kept at 4 °C for less than 24 h before the storage at − 80 °C for future usage. DNA Extraction Kit (TANTICA, Nanjing, China) was used to extract DNA from whole blood and further bisulfite-converted utilized EZ-96 DNA Methylation Gold Kit according to standard protocol (Zymo Research, Orange, U.S.). All the samples were processed in parallel.

Fang Y., Qu Y., Ji L., Sun H., Li J., Zhao Y., Liang F., Wang Z., Su J., Liu J., Dai L, & Ouyang S. (2022). Novel blood-based FUT7 DNA methylation is associated with lung cancer: especially for lung squamous cell carcinoma. Clinical Epigenetics, 14, 167.

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Whole-Genome DNA Extraction and Bisulfite Treatment

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Peripheral blood was collected using Ethylene Diamine Tetraacetic Acid (EDTA) tubes and stored at −80°C. Whole-genome DNA was extracted from the whole blood sample according to the instructions of the DNA extraction kit (TANTICA, China). The above-described extracted DNA was treated with bisulfite using the EZ-96 DNA Methylation Gold Kit (Zymo Research, USA) kit according to the above instructions.

Li J., Zhou X., Li L., Ji L., Li J., Qu Y., Wang Z., Zhao Y., Zhang J., Liang F., Liu J., Gu W., Yang R., Ma F, & Dai L. (2023). The association between CTSZ methylation in peripheral blood and breast cancer in Chinese women. Frontiers in Oncology, 13, 1148635.

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Bisulfite Conversion and Methylation Profiling

Cited 1 time

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Bisulfite conversion of 500 ng DNA per sample was conducted using the EZ-96 DNA Methylation Gold Kit (Zymo Research, Irvine, CA). Samples subsequently were processed for Illumina (San Diego, CA) HumanMethylation450 BeadChips and Infinium MethylationEPIC arrays according to the manufacturers’ instruction, and specimens were randomized across arrays. Methylation status was reported as the β value (range, 0–1, with 0 = unmethylated and 1 = fully methylated). For differential methylation analysis and principle component analysis the M value was used.⁴⁶ (link)

Lewis S.K., Nachun D., Martin M.G., Horvath S., Coppola G, & Jones D.L. (2019). DNA Methylation Analysis Validates Organoids as a Viable Model for Studying Human Intestinal Aging. Cellular and Molecular Gastroenterology and Hepatology, 9(3), 527-541.

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DNA Extraction and Bisulfite Treatment

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The DNeasy Blood and Tissue Kit (Qiagen) and the phenol-chloroform method were used for DNA extraction from frozen tumors, cell lines, and FFPE samples. After that, sodium bisulfite treatment ranging from 500–1000 ng of genomic DNA was performed using EZ-96 DNA Methylation-Gold Kit (Zymo Research, Irvine, CA, USA), according to the manufacturer’s protocol.

Lobo J., Nunes S.P., Gillis A.J., Barros-Silva D., Miranda-Gonçalves V., van den Berg A., Cantante M., Guimarães R., Henrique R., Jerónimo C, & Looijenga L.H. (2019). XIST-Promoter Demethylation as Tissue Biomarker for Testicular Germ Cell Tumors and Spermatogenesis Quality. Cancers, 11(9), 1385.

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DNA Extraction and Bisulfite Conversion

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DNA was extracted from the frozen tissue and urine samples using the AllPrep DNA Mini kit (Qiagen, Inc., Valencia, CA, USA) according to the manufacturer's protocol. The concentration and purity of DNA were assessed by determining the OD₂₆₀/OD₂₈₀ ratio using a Nanodrop 1000 spectrophotometer (Thermo Fisher Scientific, Inc., Waltham, MA, USA). DNA was treated by bisulfite using an EZ-96 DNA Methylation-Gold™ kit (Zymo Research, Irvine, CA, USA) according to the manufacturer's protocol, as well as the previously described protocol (20 (link)), and stored at −80°C until use.

Xin J., Xu R., Lin S., Xin M., Cai W., Zhou J., Fu C., Zhen G., Lai J., Li Y, & Zhang P. (2016). Clinical potential of TCF21 methylation in the diagnosis of renal cell carcinoma. Oncology Letters, 12(2), 1265-1270.

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Methylation Analysis of Sperm DNA

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Extracted sperm DNA was bisulfite converted with EZ-96 DNA Methylation-Gold kit (Zymo Research, Irvine CA) according to the manufacturer’s recommendations specifically for use with array platforms. Converted DNA was then delivered to the University of Utah Genomics Core Facility hybridized to Infinium HumanMethylation450 BeadChip microarrays (Illumina) and analyzed according to Illumina protocols. Array data for all samples were evaluated for standard data quality indicators, and two samples (both general population samples, one smoker and one non-smoker) were excluded from the study due to failure to meet established standards.

Jenkins T., James E., Alonso D., Hoidal J., Murphy P., Hotaling J., Cairns B., Carrell D, & Aston K. (2017). Cigarette Smoking Significantly Alters Sperm DNA Methylation Patterns. Andrology, 5(6), 1089-1099.

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