Qpcr core kit for sybr green 1 no rox

Manufactured by Eurogentec

Sourced in Belgium

The QPCR Core kit for SYBR Green I No ROX is a laboratory equipment product designed for quantitative real-time PCR (qPCR) applications. The kit includes all the necessary components, such as the SYBR Green I dye and PCR master mix, to perform quantitative PCR reactions without the need for a reference dye.

Automatically generated - may contain errors

Lab products found in correlation

Trizol, by Thermo Fisher Scientific (2 mentions) Myiq single color real time pcr detection system, by Bio-Rad (2 mentions) Revertaid rt kit, by Thermo Fisher Scientific (2 mentions) Turbo dna free kit, by Thermo Fisher Scientific (2 mentions) Tripure, by Roche (2 mentions) Nucleospin rna plant, by Macherey-Nagel (1 mentions) Quick amp labeling kit, by Agilent Technologies (1 mentions) Rnaeasy plant mini kit, by Qiagen (1 mentions) Mx3005p qpcr machine, by Eurogentec (1 mentions) Primescript rt qpcr kit, by Takara Bio (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Iscript gdna clear cdna synthesis kit, by Bio-Rad (1 mentions) Cfx96 touch real time pcr detection system, by Bio-Rad (1 mentions) Revertaid reverse transcriptase, by Thermo Fisher Scientific (1 mentions) Sybr green reaction mix, by Eurogentec (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Lightcycler 480, by Roche (1 mentions) Iscript reverse transcription kit, by Bio-Rad (1 mentions) Rnase free dnase, by Qiagen (1 mentions) Pegfp c1, by Promega (1 mentions)

Spelling variants of this product

SYBR Green (34 citations) QPCR Core kit for SYBR Green I (27 citations) QPCR Core Kit (14 citations) SYBR Green I (10 citations) SYBR Green I Core Kit (8 citations) QPCR Core kit for SYBR Green (6 citations) QPCR SYBR Green Core Kit (4 citations) SYBR Green qPCR kit (4 citations) QPCR kit for SYBR Green I (3 citations) SYBR® Green kit (2 citations)

8 protocols using qpcr core kit for sybr green 1 no rox

Transcript Abundance Analysis Protocol

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

For transcript abundance analysis, RNA was extracted from the indicated tissues using the RNAeasy Plant Mini Kit (Qiagen) or NucleoSpin RNA Plant (Macherey-Nagel) according to the manufacturer’s instructions. Isolated RNA was used for microarray or reverse transcription. cDNA was prepared by using the PrimeScript RT-qPCR Kit (TaKaRa). For microarray analysis, extracted RNA was labeled and hybridized according to the protocol of the Quick Amp labeling kit (Agilent, http://www.agilent.com/home). Agilent single-color technology arrays (60 k) were used for the hybridizations. Raw intensity data were normalized with the quantile method, and subsequently, the low expression probes were discarded after log2 transformation. Differentially expressed probes were filtered after pairwise comparisons (|log2FC| ≥ 1, FDR ≤ 0.05).
RT-qPCR was performed on a Stratagene Mx3005P qPCR machine using a SYBR Green containing reaction mix (Eurogentec, qPCR Core kit for SYBR Green I No ROX). For all qPCR analysis, N. attenuata IF-5A housekeeping gene was used as internal reference. Primer sequences were listed in the S2 Table.

Li S., Joo Y., Cao D., Li R., Lee G., Halitschke R., Baldwin G., Baldwin I.T, & Wang M. (2020). Strigolactone signaling regulates specialized metabolism in tobacco stems and interactions with stem-feeding herbivores. PLoS Biology, 18(8), e3000830.

+ Open protocol

+ Expand

RT-qPCR Analysis of Transgene Expression

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

HEK293 GLA^−/− cells were grown in 12-well plates in complete medium and transiently transfected using Lipofectamine 2000 (Life Technologies) according to the manufacturer's protocol. RNA was extracted 72 h after transfection by using PRImeZOL ™ reagent following the manufacturer's protocol. RNA was reverse-transcribed using iScript™ gDNA Clear cDNA Synthesis Kit (Bio-Rad, Hercules, CA, USA) following the manufacturer's protocol. Real-time RT-qPCR was performed on the CFX96 Touch Real-Time PCR Detection System (Bio-RAD) using the qPCR Core kit for SYBR® Green I No ROX (Eurogentec, Liège, Belgium) with specific primers for the indicated genes (pGLA refers to transfected GLA).

Unlabelled Table

	Forward	Reverse
HSPA5	5’-CGCTGAGGCTTATTTGGGAAA-3′	5’-TGCCGTAGGCTCGTTGATG −3’
XBP1S	5’-GAGTCCGCAGCAGGTG-3′	5’-ATACCGCCAGAATCCATGG-3’
DDIT3	5’-GCTGGAACCTGAGGAGAGAG-3′	5’-TTCTTCCTCTTCATTTCCAG-3′
pGLA	5’-ACAGCTCCTCCCTGTGAAAAG-3’	5’-GATTACGCTTGATAAACCCGCTG-3’
HPRT1	5’-AGCCCTGGCGTCGTGATTAGT-3’	5’-TGTGATGGCCTCCCATCTCC-3’

Consolato F., De Fusco M., Schaeffer C., Pieruzzi F., Scolari F., Gallieni M., Lanzani C., Feriozzi S, & Rampoldi L. (2022). α-Gal A missense variants associated with Fabry disease can lead to ER stress and induction of the unfolded protein response. Molecular Genetics and Metabolism Reports, 33, 100926.

+ Open protocol

+ Expand

Transcriptome Analysis by qPCR

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

RNA extraction was performed with TRIzol (Invitrogen, Darmstadt, Germany), according to the manufacturer's instructions. cDNA was synthesized by reverse transcription using oligo(dT) primer and RevertAid reverse transcriptase (Invitrogen). qPCR was performed using actin as standard on a Stratagene Mx3005P qPCR machine using a SYBR Green containing reaction mix (Eurogentec, Cologne, D, http://www.eurogentec.com/; qPCR Core kit for SYBR Green I No ROX). The primer sequences are provided in Table S2.

Schäfer M., Meza-Canales I.D., Navarro-Quezada A., Brütting C., Vanková R., Baldwin I.T, & Meldau S. (2014). Cytokinin levels and signaling respond to wounding and the perception of herbivore elicitors in Nicotiana attenuata. Journal of Integrative Plant Biology, 57(2), 198-212.

+ Open protocol

+ Expand

RNA Extraction, cDNA Synthesis, and qPCR Analysis

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

RNA was extracted with TRIzol (Invitrogene), according to the manufacturer instructions. cDNA was synthesized by reverse transcription using oligo(dT) primer and RevertAid reverse transcriptase (Invitrogen). qPCR was performed using actin as standard on a Stratagene Mx3005P qPCR machine using a SYBR Green reaction mix (Eurogentec; qPCR Core kit for SYBR Green I No ROX). The primer sequences are provided in Table S16.

Brütting C., Schäfer M., Vanková R., Gase K., Baldwin I.T, & Meldau S. (2017). Changes in cytokinins are sufficient to alter developmental patterns of defense metabolites in Nicotiana attenuata. The Plant journal : for cell and molecular biology, 89(1), 15-30.

+ Open protocol

+ Expand

Quantification of yedYZ mRNA Expression

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

Expression levels of the yedYZ (msrPQ) mRNA were assessed in M63 minimal medium supplemented with 0.5% glycerol, 0.15% casaminoacids, 1 mM MgSO₄, 1 mM MoNa₂O₄, 17 μM Fe₂(SO₄)₃, and vitamins (thiamin 10 μg/ml, biotin 1 μg/ml, riboflavin 10 μg/ml, and nicotinamide 10 μg/ml). Overnight cultures of MG1655 were diluted to an A₆₀₀ of 0.04 in fresh M63 minimal medium (100 ml) and cultured aerobically at 37°C until an A₆₀₀ of 0.8. Cells (10 ml) were then pelleted, resuspended in TriPure (Roche) and homogenized. After mixing with chloroform, RNA was isolated by centrifugation (15 min, 15,700 × g, 4°C), precipitated with isopropanol, washed with ethanol 70%, dried and finally resuspended in DEPC water. Any residual DNA was eliminated by treatment of the sample with DNAse (Turbo DNA-free^™ Kit, Ambion). The RevertAid RT kit (Thermo Scientific) was used to generate cDNA from 1 μg RNA extracted from each of the cultured strains. cDNAs were then diluted 1/10 and submitted to real time PCR (qPCR), using the qPCR Core kit for SYBR Green I No ROX (Eurogentec) and a MyiQ^™ Single-Color Real-Time PCR Detection System (Bio-Rad). yedYZ expression levels were normalized to the expression of gapA. Primers used for qPCR analysis were: qPCR_yedYZ_Fw and qPCR_yedYZ_Rv for yedYZ, and qPCR_gapA_Fw and qPCR_gapA_Rv for gapA (Supplementary Table 3).

Gennaris A., Ezraty B., Henry C., Agrebi R., Vergnes A., Oheix E., Bos J., Leverrier P., Espinosa L., Szewczyk J., Vertommen D., Iranzo O., Collet J.F, & Barras F. (2015). Repairing oxidized proteins in the bacterial envelope using respiratory chain electrons. Nature, 528(7582), 409-412.

+ Open protocol

+ Expand

RNA Extraction and qPCR Analysis

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

RNA was extracted from confluent mTAL or MDCK cells grown in 35 mm dishes using 1 mL Isol-RNA Lysis Reagent (5Prime) following the manufacturer protocol. Extracted RNA was treated with RNase-free DNase (Qiagen) and cleaned-up using the RNeasy Mini kit (Qiagen). RNA was reverse-transcribed using the iScript Reverse Transcription kit (Bio-Rad, Hercules, CA). Real-time RT-qPCR was performed on the LightCycler 480 instrument (Roche, Basel, Switzerland) using the qPCR Core kit for SYBR® Green I No ROX (Eurogentec, Liège, Belgium) with specific primers for the indicated genes (Table 2).

Schaeffer C., Merella S., Pasqualetto E., Lazarevic D, & Rampoldi L. (2017). Mutant uromodulin expression leads to altered homeostasis of the endoplasmic reticulum and activates the unfolded protein response. PLoS ONE, 12(4), e0175970.

+ Open protocol

+ Expand

Quantification of yedYZ mRNA Expression

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

+ Open protocol

+ Expand

Quantitative Analysis of Genomic DNA

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

Genomic DNA was extracted from cells by proteinase K digestion and phenol:chloroform extraction followed by ethanol precipitation and resuspension overnight in TE (pH 8.0). Genomic DNA concentration was quantified with UV spectrophotometer (NanoDrop) and diluted to 16 ng/μl for qPCR. For standard curve, a series of mixtures in which the number of pEGFP-C1 (Promega) plasmid molecules ranged from 128 to 1024 copies per HeLa genome were prepared using HeLa genomic DNA. 32 ng of genomic DNA from different cell lines or copy number standards were subjected to quantitative PCR. Quantitative PCR was performed on Roche LightCycler 2.0 using qPCR Core kit for SYBR® Green I No ROX (Eurogentec). qPCR reactions with copy number standards were performed in duplicate. qPCR reactions with cell line genomic DNAs were performed in triplicate. Melting curve analysis was carried out at the end of cycling to confirm amplification of a single PCR product. Following EGFP and human TRKB (genomic control) specific PCR primer sets were used: EGFP sense 5′- CAG AAG AAC GGC ATC AAG GTG-3′, antisense 5′- TGG GTG CTC AGG TAG TGG TTG -3′; TRKB sense 5′- CAC AGG GCT CCT TAA GGA TAA C -3′, antisense 5′- GCA CAG TGA GGT TGA CAG AAT C-3′. Copy number estimates were calculated with qBASEPlus 2.6 software (Biogazelle) using EGFP as target and TRKB as reference.

Jaanson K., Sepp M., Aid-Pavlidis T, & Timmusk T. (2014). BAC-based cellular model for screening regulators of BDNF gene transcription. BMC Neuroscience, 15, 75.

+ 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!