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Gotaq sybr green

Manufactured by Promega
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GoTaq SYBR green is a DNA polymerase enzyme used for real-time PCR amplification. It contains SYBR Green I dye for detecting and quantifying DNA during the amplification process.

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Lab products found in correlation

Random primer, by New England Biolabs (2 mentions) Trizol, by Thermo Fisher Scientific (2 mentions) Cfx96 touch deep well rt pcr system, by Bio-Rad (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Reverse transcription kit, by Promega (1 mentions) M mulv reverse transcriptase, by Promega (1 mentions) Quick rna kit, by Zymo Research (1 mentions) Real time pcr system, by Bio-Rad (1 mentions) Rt2 first strand kit, by Qiagen (1 mentions) Rt2 profiler, by Qiagen (1 mentions) Rneasy plus kit, by Qiagen (1 mentions)

4 protocols using gotaq sybr green

1

Quantitative Analysis of Circular RNA Expression

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Total RNA was extracted from tissues and cell samples with TRIzol Reagent (Invitrogen, Carlsbad, CA). RNA concentration was detected using a Thermo Nanodrop 2000 spectrophotometer (Waltham, MA). RNA quality was evaluated according to the A260/A280 ratio. RNA was reverse transcribed to produce cDNA with a Reverse Transcription Kit (Promega, Madison, WI). RT-qPCR was performed using GoTaq SYBR Green (Promega, Madison, WI) with a CFX96 Touch Deep Well RT-PCR System (Bio-Rad, Hercules, CA). The specific primers are as follows: β-actin (forward: 5ʹ-CTCTGCCCGCATGAACCT-3ʹ, reverse: 5ʹ-CCACCATCCACATCCCAC-3ʹ) and circ-1073 (forward: 5ʹ-AGTCAGTTCCTTGTGGAGCC-3ʹ, reverse: 5ʹ-GCATGGGTTCTGACGGACAT-3ʹ). β-Actin was amplified as the reference standard for normalization.
Yi Z., Li Y., Wu Y., Zeng B., Li H., Ren G, & Wang X. (2020). Circular RNA 0001073 Attenuates Malignant Biological Behaviours in Breast Cancer Cell and Is Delivered by Nanoparticles to Inhibit Mice Tumour Growth. OncoTargets and therapy, 13, 6157-6169.
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2

Quantifying mRNA Levels by RT-qPCR

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Total RNA was extracted from cells using the Quick-RNA kit (Zymo) according to manufacturer’s protocol. RNA was used to generate cDNA by reverse transcriptase PCR using the M-MuLV Reverse Transcriptase (Promega). Relative mRNA expression levels were determined using the Go-Taq SYBR Green (Promega) on a Bio-Rad Real-Time PCR System with selected primer pairs. Expression levels were normalized to Gapdh, used as a control housekeeping gene, and computed as described previously (Ferrari et al., 2014 (link)).
Scelfo A., Fernández-Pérez D., Tamburri S., Zanotti M., Lavarone E., Soldi M., Bonaldi T., Ferrari K.J, & Pasini D. (2019). Functional Landscape of PCGF Proteins Reveals Both RING1A/B-Dependent-and RING1A/B-Independent-Specific Activities. Molecular Cell, 74(5), 1037-1052.e7.
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3

Analyzing WNT Signaling in Muscle Cells

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RNA was prepared from cells or from whole muscle tissue (after grinding with plastic pestles), using Trizol (Invitrogen); after DNAse treatment, cDNA was synthesized using MMuLV reverse transcriptase and random primers (NEB). Real time RT-PCR was performed using a Corbett Rotogene and GoTaq SYBR green (Promega). Primers are listed in Supplementary Table S3. For PCR arrays, RNA was prepared from pooled primary myoblast cultures using the RNeasyPlus Kit (Qiagen). cDNA was synthesized using RT2 First Strand Kit (SABiosciences) and applied to RT2 Profiler™ Mouse WNT Signaling Targets (PAMM-243A) and Wnt Signaling Pathway (PAMM-043A) arrays (these arrays share some genes). Analysis used an Applied Biosystems 7300 Real-Time PCR System and SABiosciences online software to calculate fold-change and p-value (each sample was run three times for each array).
Zhuang L., Hulin J.A., Gromova A., Nguyen T.D., Yu R.T., Liddle C., Downes M., Evans R.M., Makarenkova H, & Meech R. (2014). Barx2 and Pax7 have antagonistic functions in regulation of Wnt signaling and satellite cell differentiation. Stem cells (Dayton, Ohio), 32(6), 1661-1673.
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4

Quantitative RT-PCR Analysis of Gene Expression

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RNA was prepared from cells using TRIzol (Life Technologies, Carlsbad, California, www.lifetechnologies.com); after DNase treatment, cDNA was synthesized using NxGen M-MuLV reverse transcriptase (Lucigen, Wisconsin, www.lucigen.com) and random primers (New England Biolabs, Ipswich, Massachusetts, www.neb.com). Quantitative RT-PCR was performed using a Corbett Rotorgene (Qiagen, Venlo, Limburg, Netherlands, www.qiagen.com) and GoTaq SYBR green (Promega). Significance was assessed using Student’s t test.
Hulin J.A., Nguyen T.D., Cui S., Marri S., Yu R.T., Downes M., Evans R.M., Makarenkova H, & Meech R. (2016). Barx2 and Pax7 regulate Axin2 expression in myoblasts by interaction with β-catenin and chromatin remodelling. Stem cells (Dayton, Ohio), 34(8), 2169-2182.
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