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Epitaq polymerase

Manufactured by Takara Bio
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Sourced in China, Japan

EpiTaq polymerase is a high-fidelity DNA polymerase designed for accurate DNA amplification. It exhibits proofreading activity, ensuring precise replication of DNA sequences.

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Lab products found in correlation

Ez dna methylation kit, by Zymo Research (2 mentions) Pgem t vector, by Promega (2 mentions) Topo ta cloning kit, by Thermo Fisher Scientific (1 mentions) Quick dna miniprep kit, by Zymo Research (1 mentions) Ez dna methylation gold kit, by Zymo Research (1 mentions) Epimark bisulfite conversion kit, by New England Biolabs (1 mentions) Wizard plus sv minipreps dna purification system, by Promega (1 mentions) E coli jm109 competent cells, by Promega (1 mentions) Pgem t easy vector, by Promega (1 mentions)

4 protocols using epitaq polymerase

1

Bisulfite Sequencing for DNA Methylation Analysis

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Bisulfite sequencing was performed to validate the DMRs detected by the whole-genome DNA methylation analysis. DNA (330 ng) was isolated using Quick-DNA Miniprep Kit (Zymo Research, Orange, CA, USA) and DNA bisulfite conversion was conducted by using the EZ DNA Methylation Gold Kit (Zymo Research, Orange, CA, USA). Bisulfite-converted DNA (1 µg) was amplified by PCR with EpiTaq polymerase (for bisulfite-treated DNA) (TaKaRa Biotechnology Co., Ltd., Dalian, China), and primers (Supplementary File 1) were manually designed using Primer Premier 5. Before primer design, DNA sequences need to be methylation converted in the MethPrimer website (http://www.urogene.org/methprimer/). PCR products were cloned using the Topo TA cloning kit (Invitrogen Corporation, Carlsbad, CA, USA) and after verification of successful cloning, at least ten different clones for each sample were sent for sequencing at TSINGKE Biological Technology. The DNA methylation profile of individual CpG sites were analyzed and presented using BiQ Analyzer (https://biq-analyzer.bioinf.mpi-inf.mpg.de/).
Guo L., Cheng H., Fu S., Liu J., Zhang Y., Qiu Y, & Chen H. (2022). Methylome and Transcriptome-Based Integration Analysis Identified Molecular Signatures Associated With Meningitis Induced by Glaesserella parasuis. Frontiers in Immunology, 13, 840399.
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2

Bisulfite Conversion and Sequencing

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Bisulfite conversion of 400 ng of genomic DNA was performed using the EZ DNA Methylation Kit (Zymo Research, D5001). The investigated sequences were amplified by PCR using EpiTaq polymerase (Takara Bio, R110B) and primers described in Supplementary Data 1. PCR products were cloned into the pGEM-T vector (Promega, A1360) and sequenced with the M13 reverse primer.
Pachano T., Sánchez-Gaya V., Ealo T., Mariner-Faulí M., Bleckwehl T., Asenjo H.G., Respuela P., Cruz-Molina S., Martín M.M., Haro E., van IJcken W.F., Landeira D, & Rada-Iglesias A. (2021). Orphan CpG islands amplify poised enhancer regulatory activity and determine target gene responsiveness. Nature genetics, 53(7), 1036-1049.
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3

Bisulfite Conversion and Sequencing

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Bisulfite conversion of 400 ng of genomic DNA was performed using the EZ DNA Methylation Kit (Zymo Research, D5001). The investigated sequences were amplified by PCR using EpiTaq polymerase (Takara Bio, R110B) and primers described in Supplementary Data 1. PCR products were cloned into the pGEM-T vector (Promega, A1360) and sequenced with the M13 reverse primer.
Pachano T., Sánchez-Gaya V., Ealo T., Mariner-Faulí M., Bleckwehl T., Asenjo H.G., Respuela P., Cruz-Molina S., Martín M.M., Haro E., van IJcken W.F., Landeira D, & Rada-Iglesias A. (2021). Orphan CpG islands amplify poised enhancer regulatory activity and determine target gene responsiveness. Nature genetics, 53(7), 1036-1049.
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4

Bisulfite Sequencing of Fetal DNA

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Purified genomic DNA derived from the fetus used for allelic expression analysis was treated with sodium bisulphite solution using EpiMark Bisulfite Conversion kit (New England Biolabs, Massachusetts, USA). After the bisulphite treatment of the genomic DNA, 40 cycles of PCR were carried out using EpiTaq polymerase (Takara Bio, Shiga, Japan) with the primers (Additional file 1) designed by MethPrimer (https://www.urogene.org/methprimer/) [66 (link)]. The PCR products were cloned using a pGEM T-easy vector (Promega, Wisconsin, USA) and E. coli JM109 competent cells (Promega, Wisconsin, USA). Plasmids were purified using Wizard Plus SV Minipreps DNA Purification System (Promega, Wisconsin, USA) and directly sequenced using M13 primers (Additional file 1). The sequence data were analysed by quantification tool for methylation analysis (QUMA) programme (http://quma.cdb.riken.jp) [67 (link)].
Ishihara T., Griffith O.W., Suzuki S, & Renfree M.B. (2022). Placental imprinting of SLC22A3 in the IGF2R imprinted domain is conserved in therian mammals. Epigenetics & Chromatin, 15, 32.
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