Power sybr green

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom, Germany, Canada, Switzerland, Australia

Power SYBR Green is a fluorescent dye used for quantitative real-time PCR (qPCR) applications. It binds to double-stranded DNA and emits a fluorescent signal upon excitation, allowing the quantification of DNA during the PCR amplification process.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Power SYBRs Green PCR Master Mix (10 citations) Power SYBR Green RT-PCR kit (20 citations) Power tracker SYBR Green Master Mix (2 citations) Power SYBR Green PCR 2 X Master Mix (2 citations) Power SYBR Green Mater Mix (3 citations) Fast Power SYBR Green PCR master mix (3 citations) Power SYBR Green PCR Master Mix and RT-PCR (2 citations) Power-up Sybr Green 2 × Master Mix (5 citations) Power SYBR Green PCR Master Mix 2x (16 citations) Power SYBR Green 2 × Mastermix (3 citations)

542 protocols using power sybr green

Oxtr Gene Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Extraction of RNA was done using the Qiagen AllPrep DNA/RNA Mini Kit (Qiagen, Valencia, CA) following manufacturer instructions. RNA was processed for cDNA synthesis following the protocol provided in the iScript cDNA Synthesis kit (Bio-Rad, Hercules, CA). Real-time PCR for the Oxtr main transcript was conducted using a 7500 Fast Real-Time PCR System (Applied Biosystems) using Power SYBR Green (Applied Biosystems No. 4367659). Real-time PCR for the Oxtr alternative transcript was conducted using the CFX96 System (Bio-Rad) using Power SYBR Green (Applied Biosystems). Real-time PCR for Pgk1 was completed on both Real-time PCR systems. See Additional file 2 for all RT-PCR primers and cycling conditions. All reactions were run in triplicate (replicate standard deviation was < 0.05) and their specificity verified by melting curve analysis and separation on a 2% agarose gel. Primer performance was evaluated using standard serial dilution and all primer sets performed within acceptable range for efficiency (See Additional file 2). Relative gene expression is presented using the comparative Ct method, 2^−ΔCt, comparing target expression to Pgk1 expression measured on the same real-time PCR system. Pgk1 was chosen as a reference based on data in mouse brain showing its reliability across brain regions and developmental time points [58 (link)].

Danoff J.S., Wroblewski K.L., Graves A.J., Quinn G.C., Perkeybile A.M., Kenkel W.M., Lillard T.S., Parikh H.I., Golino H.F., Gregory S.G., Carter C.S., Bales K.L, & Connelly J.J. (2021). Genetic, epigenetic, and environmental factors controlling oxytocin receptor gene expression. Clinical Epigenetics, 13, 23.

+ Open protocol

+ Expand

Comprehensive RNA Extraction and qPCR Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA extraction was performed using TRI-reagent (Sigma-Aldrich, Merck Millipore, Guyancourt, France) to isolate the aqueous fraction, followed by column-based extraction (ZymoResearch, distributed by Ozyme, Saint-Cyr-l’École, France). The RNA concentration was measured via NanoDrop 2000 (Thermo Fisher Scientific, Illkirch-Graffenstaden, France). The cDNA strand was synthesized from 500 ng of total RNA (Thermo Fisher Scientific). Quantification of KRAS, NRAS, and RPLP0 genes was measured via Power-Sybr-Green assays (Thermo Fisher Scientific) with the StepOne™ Real-time PCR System in agreement with The Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines [21 (link)]. The qPCR primers are referenced in the Supplementary Table S1.

Vidal-Cruchez O., Nicolini V.J., Rete T., Jacquet K., Rezzonico R., Lacoux C., Domdom M.A., Roméo B., Roux J., Hubstenberger A., Mari B., Mograbi B., Hofman P, & Brest P. (2023). KRAS and NRAS Translation Is Increased upon MEK Inhibitors-Induced Processing Bodies Dissolution. Cancers, 15(12), 3078.

+ Open protocol

+ Expand

Quantifying Primary Transcript Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols

We prepared RNA using TRIzol™ (ThermFisher Scientific, Cat # 15596018) and purified using the RNeasy Mini kit with on-column DNase treatment (QIAGEN, Cat # 74106). Reverse transcriptase reactions were performed using iScript (Bio-Rad, Cat #1708841) and qPCR reactions were prepared with Power SYBR Green (ThermoFisher Scientific). All primers flank intron exon-junctions, and primer sets are listed in Table S2. Primary transcript quantities are first normalized to Pabpc1 mature mRNA, similar results were obtained when normalizing to Gapdh mature mRNA (data not shown). Following this, transcript levels at each time point were additionally normalized to the mean transcript quantity across all time points, so that the activation kinetics for different primary transcripts could be compared.

Behera V., Stonestrom A.J., Hamagami N., Hsiung C.C., Keller C.A., Giardine B., Sidoli S., Yuan Z.F., Bhanu N.V., Werner M.T., Wang H., Garcia B.A., Hardison R.C, & Blobel G.A. (2019). Interrogating Histone Acetylation and BRD4 as Mitotic Bookmarks of Transcription. Cell reports, 27(2), 400-415.e5.

+ Open protocol

+ Expand

Gene Expression Analysis of 3D and 2D Cell Cultures

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was extracted from 3D cultures or from 2D monolayer cells using Trizol (Thermo Fisher Scientific). Complementary DNA was synthesized with a SuperScript III First-Strand Synthesis System (Thermo Fisher Scientific) according to the manufacturer’s instructions. qPCR was performed in duplicate for each gene on a StepOnePlus Real-Time PCR System (Thermo Fisher Scientific) using Power SYBR Green (Thermo Fisher Scientific). RPS18 was used as an internal control. Fold difference in gene expression was determined by the 2-ΔΔCt method. The sequence of the primers for qPCR are shown in Supplementary Table S1 online.

Kawai S., Nakano K., Tamai K., Fujii E., Yamada M., Komoda H., Sakumoto H., Natori O, & Suzuki M. (2021). Generation of a lung squamous cell carcinoma three-dimensional culture model with keratinizing structures. Scientific Reports, 11, 24305.

+ Open protocol

+ Expand

Quantitative Gene Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

To analyze gene expression, cells were lysed and RNA purified using the RNeasy Mini Kit (Qiagen). To generate cDNA, a total of 1 μg RNA was used for reverse transcription using the High Capacity cDNA Reverse Transcription Kit (ThermoFisher Scientific). Using a Stratagene Mx3000P RT-PCR machine. RT-qPCR was performed using either TaqMan probes with the TaqMan Advanced 2x Master mix (ThermoFisher Scientific) or primers with Power SybrGreen (ThermoFisher Scientific). All TaqMan probes were run with a cycle of 15s at 95°C and 30s at 60°C for 40 cycles. All primers were used at 1 μM and run with cycles of 15s at 95°C and 45s at 60°C for 40 cycles. Taqman probes and primers used are listed below.
TaqMan probes used in this study:
Primers for RT-qPCR using SybrGreen:

Reeves A.R., Spiller K.L., Freytes D.O., Vunjak-Novakovic G, & Kaplan D.L. (2015). Controlled Release of Cytokines Using Silk-biomaterials for Macrophage Polarization. Biomaterials, 73, 272-283.

+ Open protocol

+ Expand

Quantitative RNA Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were grown and collected at log-phase growth. RNA was extracted using RNeasy Mini Kit (Qiagen, Germantown, MD, USA). RNA was normalized using Nanodrop 2000 (ThermoFisher Scientific) and cDNA libraries were created with High-Capacity cDNA Reverse Transcription Kit (ThermoFisher Scientific). cDNA libraries were amplified with Power SYBR Green on a QuantStudio6 with optimized primers for various fusions (Supplementary Table 12 for primers used in this study; ThermoFisher Scientific).

Hong A.L., Tseng Y.Y., Cowley G.S., Jonas O., Cheah J.H., Kynnap B.D., Doshi M.B., Oh C., Meyer S.C., Church A.J., Gill S., Bielski C.M., Keskula P., Imamovic A., Howell S., Kryukov G.V., Clemons P.A., Tsherniak A., Vazquez F., Crompton B.D., Shamji A.F., Rodriguez-Galindo C., Janeway K.A., Roberts C.W., Stegmaier K., van Hummelen P., Cima M.J., Langer R.S., Garraway L.A., Schreiber S.L., Root D.E., Hahn W.C, & Boehm J.S. (2016). Integrated genetic and pharmacologic interrogation of rare cancers. Nature Communications, 7, 11987.

+ Open protocol

+ Expand

Quantifying Gene Expression in A. woodii

Check if the same lab product or an alternative is used in the 5 most similar protocols

Complementary DNA was synthesized from 1 µg of RNA using the Omniscript RT Kit (QIAGEN), random hexamer (Thermo Fisher), and RNase inhibitor (NEB) following the manufacturer’s instructions.
Power SYBR Green (Thermo Fisher) was used for the qPCR, and primer pairs targeting the gyrA gyrase gene in the A. woodii genome, orfB of the plasmid backbones, and the FAST gene employed are listed in Supplementary Table S8. Standard curves allowed for efficiencies of primer pairs to be calculated. Gene expression was calculated for each sample using the Pfaffl method without a calibrator gene:

E x p r e s s i o n = \frac{{E_{t a r g e t}}^{- {C p}_{t a r g e t}}}{{E_{r e f e r e n c e}}^{- {C p}_{r e f e r e n c e}}} .

Poulalier-Delavelle M., Baker J.P., Millard J., Winzer K, & Minton N.P. (2023). Endogenous CRISPR/Cas systems for genome engineering in the acetogens Acetobacterium woodii and Clostridium autoethanogenum. Frontiers in Bioengineering and Biotechnology, 11, 1213236.

+ Open protocol

+ Expand

Quantitative RT-PCR Gene Expression Analysis

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

qRT-PCR using power SYBR green (Thermo Fisher Scientific) was performed as previously described (Smith et al., 2013 (link)) using an Applied Biosystems 7500 Fast Real-Time PCR System. All primers (Table 1) were tested for efficiency using serial dilutions, and results were normalized to 18S RNA levels; data analyses were performed using the standard curve analysis method (Rutledge and Côté, 2003 (link); Smith et al., 2014 (link)). The 18S ribosomal transcript was used as a housekeeping gene as our hypoxia treatments did not alter its expression (Table 2).

Kiernan E.A., Ewald A.C., Ouellette J.N., Wang T., Agbeh A., Knutson A.O., Roopra A.S, & Watters J.J. (2020). Prior Hypoxia Exposure Enhances Murine Microglial Inflammatory Gene Expression in vitro Without Concomitant H3K4me3 Enrichment. Frontiers in Cellular Neuroscience, 14, 535549.

+ Open protocol

+ Expand

Quantitative Gene Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was extracted using the GeneJET purification kit (K0732, Thermo Scientific) and 2μg of RNA was used to
synthesize cDNA with the High-Capacity cDNA Reverse Transcription kit (4368814, Applied Biosystems™, Thermo Scientific).
The PCR reactions were performed in triplicate on the StepOnePlus system (Applied Biosystem) using Power SYBR Green (4367659,
ThermoFisher). Primer efficiency was determined and relative gene expression was measured using the primers listed in Table 2.

Marchand B., Pitarresi J.R., Reichert M., Suzuki K., Laczkó D, & Rustgi A.K. (2019). PRRX1 isoforms cooperate with FOXM1 to regulate the DNA damage response in pancreatic cancer cells. Oncogene, 38(22), 4325-4339.

+ Open protocol

+ Expand

Quantifying Primary Transcript Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!