Amplitaq dna polymerase kit

Manufactured by Thermo Fisher Scientific

Sourced in United States

AmpliTaq® DNA Polymerase Kit is a molecular biology reagent used for DNA amplification. It contains the AmpliTaq® DNA Polymerase enzyme, which catalyzes the replication of DNA strands during the polymerase chain reaction (PCR) process.

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Lab products found in correlation

Topo ta cloning vector, by Thermo Fisher Scientific (1 mentions) Epitect bisulfite kit, by Qiagen (1 mentions) Wizard genomic dna purification kit, by Promega (1 mentions) Tissuelyser, by Qiagen (1 mentions) Taqman reverse transcription kit, by Thermo Fisher Scientific (1 mentions) Rnalater, by Thermo Fisher Scientific (1 mentions) Purelink micro to midi total rna purification system, by Thermo Fisher Scientific (1 mentions) Prism 7900ht sequence detection system, by Thermo Fisher Scientific (1 mentions) Paxgene rna collection tubes, by BD (1 mentions)

Spelling variants of this product

AmpliTaq DNA polymerase (99 citations) DNA polymerase (71 citations) AmpliTaq polymerase (17 citations) Amplitaq Gold DNA Polymerase kit (17 citations) AmpliTaq polymerase kit (2 citations) AmpliTaq DNA polymerase FS Dye Terminator Cycle Sequencing Ready Reaction kit (2 citations) GeneAmp PCR Reagent Kit with AmpliTaq DNA Polymerase (1 citations)

3 protocols using amplitaq dna polymerase kit

Comprehensive Methylation Analysis Workflow

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Genomic DNA from cultured cells was isolated using the Wizard Genomic DNA Purification Kit (Promega) following the manufacturer’s protocols. ~ 1 μg genomic DNA was subjected to bisulfite conversion using EpiTect Bisulfite Kit (Qiagen). The bisulfite sequencing was obtained by three methods: sequencing of individual clones, pyrosequencing and direct sequencing of the bisulfite-treated and PCR-amplified genomic DNA. PCR amplification was obtained by AmpliTaq DNA polymerase Kit (Thermo Fisher Scientific). For the Plagl1: TSS-DMR, Peg13: TSS-DMR and Igf2r: TSS-DMR, PCR products were cloned into the TOPO TA cloning vector (Invitrogen) and individual clones were isolated and sequenced (Eurofins Genomics). For the H19/Igf2:IG-DMR, Mest:TSS-DMR and Rasgrf1:TSS-DMR, DNA methylation was assessed by Sanger sequencing (Eurofins Genomics) of the bisulfite-treated and PCR-amplified genomic DNA (AmpliTaq DNA polymerase Kit, Thermo Fisher Scientific). For the Kcnq1ot1:TSS-DMR, Meg3/Dlk1:IG-DMR, Peg3:TSS-DMR, and Snrpn:TSS-DMR DNA methylation was analysed by pyrosequencing using PyroMark Q48 Autoprep sequencing machine (Qiagen). For the H19/Igf2:IG-DMR, Mest:TSS-DMR and Rasgrf1:TSS-DMR, DNA methylation was assessed after direct sequencing of the bisulfite-treated and PCR-amplified genomic DNA. All the primers are listed in Supplementary Table 6.

Acurzio B., Verma A., Polito A., Giaccari C., Cecere F., Fioriniello S., Della Ragione F., Fico A., Cerrato F., Angelini C., Feil R, & Riccio A. (2021). Zfp57 inactivation illustrates the role of ICR methylation in imprinted gene expression during neural differentiation of mouse ESCs. Scientific Reports, 11, 13802.

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Quantitative RT-PCR Analysis of SPHK1 Expression

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Quantitative real-time PCR (qt-RT-PCR) analysis was performed by a two-step process, a 15-cycle preamplification step (AmpliTaq® DNA Polymerase Kit; Applied Biosystems Inc., Foster City, CA, USA) followed by measurement of mRNA with an ABI PRISM 7900HT Sequence Detection System. For the measurement of mRNA levels SPHK1, primers were custom designed and ordered from IDT (San Diego, CA). The sequences of the primers are: forward 5’-CTTGCAGCTCTTCCGGAGTC-3’, reverse 5’-GCTCAGTGAGCATCAGCGTG-3’.

Zhang L., Wang X., Bullock A.J., Callea M., Shah H., Song J., Moreno K., Visentin B., Deutschman D., Alsop D.C., Atkins M.B., Mier J.W., Signoretti S., Bhasin M., Sabbadini R.A, & Bhatt R.S. (2015). Anti-S1P antibody as a novel therapeutic strategy for VEGFR TKI resistant renal cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 21(8), 1925-1934.

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RNA Extraction and RT-qPCR Analysis

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Biopsy samples were immediately placed in 150µl RNAlater (Life Technologies, Grand Island, NY) and stored at −70°C. After thawing, samples were homogenized using Tissuelyser (Qiagen, Hilden, Germany), and total RNA was extracted using Purelink micro to midi Total RNA Purification System (Invitrogen, Carlsbad, CA). Peripheral blood was collected in PAXgene RNA collection tubes (BD Diagnostics, Valencia, CA), stored at room temperature for 6–24 hours, then frozen at −70°C. Total RNA was by PAXgene blood miRNA kit (PreAnalytiX, QIAGEN, Hilden, Germany). 1µg of total RNA/100 µl was converted into complementary DNA (cDNA) using Taqman Reverse Transcription kit (Applied Biosystems/Life Technologies, Foster City, CA, USA).
Real-time PCR (RT-qPCR) analysis was performed by a two-step process—a 10-cycle preamplification step (AmpliTaq® DNA Polymerase Kit; Applied Biosystems/Life Technologies, Foster City, CA, USA) followed by measurement of mRNA copies with an ABI PRISM 7900HT Sequence Detection System (for details and primers see supplemental methods).

Starling R.C., Stehlik J., Baran D.A., Armstrong B., Stone J.R., Ikle D., Morrison Y., Bridges N.D., Putheti P., Strom T.B., Bhasin M., Guleria I., Chandraker A., Sayegh M., Daly K.P., Briscoe D.M, & Heeger P.S. (2015). Multicenter Analysis of Immune Biomarkers and Heart Transplant Outcomes: Results of the Clinical Trials in Organ Transplantation-05 study. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 16(1), 121-136.

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