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Primers for real time pcr

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Primers for real-time PCR are short, synthetic DNA sequences designed to initiate the amplification of a specific target sequence during the polymerase chain reaction (PCR) process. They serve as the starting point for DNA replication, enabling the detection and quantification of target nucleic acids in real-time.

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Lab products found in correlation

Quantifast sybr green rt pcr kit, by Qiagen (4 mentions) Peqlab total rna isolation kit, by Avantor (4 mentions) Cdna synthesis kit, by Thermo Fisher Scientific (4 mentions) Rest software, by Qiagen (4 mentions) Lightcycler 480, by Roche (4 mentions) Thermal cycler, by Avantor (3 mentions) Fitc dextran, by Merck Group (1 mentions) Itaq universal sybr green one step kit, by Bio-Rad (1 mentions) Transfast, by Promega (1 mentions)

5 protocols using primers for real time pcr

1

RNA Extraction and qRT-PCR Analysis

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Total RNA was isolated using the PEQLAB total RNA isolation kit (Peqlab; Erlangen, Germany) and reverse transcription was performed in a thermal cycler (Peqlab) using a cDNA synthesis kit (Applied Biosystems; Foster City, CA). mRNA levels were examined by qRT-PCR. A QuantiFast SYBR Green RT-PCR kit (Qiagen; Hilden, Germany) was used to perform real time PCR on a LightCycler 480 (Roche Diagnostics; Vienna, Austria), and data were analyzed by the REST Software (Qiagen). Relative expression of specific genes was normalized to porcine GAPDH, respectively, as a housekeeping gene. Primers for real time PCR were obtained from Invitrogen (Vienna, Austria).
Madreiter-Sokolowski C.T., Waldeck-Weiermair M., Bourguignon M.P., Villeneuve N., Gottschalk B., Klec C., Stryeck S., Radulovic S., Parichatikanond W., Frank S., Madl T., Malli R, & Graier W.F. (2018). Enhanced inter-compartmental Ca2+ flux modulates mitochondrial metabolism and apoptotic threshold during aging. Redox Biology, 20, 458-466.
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2

Comprehensive Alzheimer's Disease Protocol

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Primers for real time PCR were obtained from Invitrogen (Thermofisher Scientific, Carlsbad, CA). The anti-Aβ (clone D54D2) antibody was obtained from Cell Signaling Technology (Danvers, MA) while anti-Aβ antibody (Clone 6E10) was obtained from Biolegend (San Diego, CA). Anti-APP (Y188) antibody was purchased from Abcam (Cambridge, MA). The Aβ 1-40 and Aβ 1-42 ELISA kits were obtained from EMD Millipore (Burlington, MA). QIAzol and RNeasy mini kit for RNA isolation were purchased from Qiagen (Germantown, MA) and iTaq Universal SYBR Green One-Step kit was from Biorad (Hercules, CA). FITC-dextran was purchased from Sigma Aldrich (St. Louis, MO). Anti-oligomer (A11) and anti-fibril (OC) antibodies were a kind gift from Prof. Rakez Kayed, University of Texas Medical Branch (UTMB), Galveston, TX.
Manocha G.D., Floden A.M., Miller N.M., Smith A.J., Nagamoto-Combs K., Saito T., Saido T.C, & Combs C.K. (2019). Temporal Progression of Alzheimer’s disease in Brains and Intestines of Transgenic Mice. Neurobiology of aging, 81, 166-176.
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3

Quantifying Gene Expression in HeLa Cells

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Total RNA was isolated using the PEQLAB total RNA isolation kit (PEQLAB Biotechnologie GmbH, Erlangen, Germany), and reverse transcription was performed in a thermal cycler (PEQLAB Biotechnologie GmbH) using a cDNA synthesis kit (Applied Biosystems, Foster City, CA, USA). Expression of MCU, UCP2, UCP3, Letm1, and AKAP-RFP-CAAX in HeLa cells was examined by RT-PCR. A QuantiFast SYBR Green RT-PCR kit (Qiagen, Hilden, Germany) was used to perform real time PCR on a LightCycler 480 (Roche Diagnostics, Vienna, Austria), and data were analyzed by the REST Software (Qiagen). Relative expression of specific genes was normalized with GAPDH as a housekeeping gene. Primers for real time PCR were obtained from Invitrogen (Vienna, Austria), and their sequences (5′-3′) were:
Madreiter-Sokolowski C.T., Gottschalk B., Parichatikanond W., Eroglu E., Klec C., Waldeck-Weiermair M., Malli R, & Graier W.F. (2016). Resveratrol Specifically Kills Cancer Cells by a Devastating Increase in the Ca2+ Coupling Between the Greatly Tethered Endoplasmic Reticulum and Mitochondria. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 39(4), 1404-1420.
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4

Gene Expression Analysis in NRK Cells

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NRK cells were transfected with IP3R-3 siRNA (see below) using Transfast (Promega Corp) using manufacturer's instructions. Total RNA was isolated using the PEQLAB total RNA isolation kit (Peqlab), and reverse transcription was performed in a thermal cycler (Peqlab) using a cDNA synthesis kit (Applied Biosystems). mRNA levels were examined by qRT-PCR. A QuantiFast SYBR Green RT-PCR kit (Qiagen) was used to perform real time PCR on a LightCycler 480 (Roche Diagnostics), and data were analyzed by the REST Software (Qiagen; https://www.qiagen.com). Relative expression of specific genes was normalized to human GAPDH as a housekeeping gene. Primers for real time PCR were obtained from Invitrogen.
Held A., Lapka J., Sargeant J., Hojanazarova J., Shaheen A., Galindo S., Madreiter-Sokolowski C., Malli R., Graier W.F, & Hay J.C. (2023). Steady-state regulation of COPII-dependent secretory cargo sorting by inositol trisphosphate receptors, calcium, and penta EF hand proteins. The Journal of Biological Chemistry, 299(12), 105471.
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5

Quantifying Gene Expression in HeLa Cells

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Total RNA was isolated using the PEQLAB total RNA isolation kit (PEQLAB Biotechnologie GmbH, Erlangen, Germany), and reverse transcription was performed in a thermal cycler (PEQLAB Biotechnologie GmbH) using a cDNA synthesis kit (Applied Biosystems, Foster City, CA, USA). Expression of MCU, UCP2, UCP3, Letm1, and AKAP-RFP-CAAX in HeLa cells was examined by RT-PCR. A QuantiFast SYBR Green RT-PCR kit (Qiagen, Hilden, Germany) was used to perform real time PCR on a LightCycler 480 (Roche Diagnostics, Vienna, Austria), and data were analyzed by the REST Software (Qiagen). Relative expression of specific genes was normalized with GAPDH as a housekeeping gene. Primers for real time PCR were obtained from Invitrogen (Vienna, Austria), and their sequences (5′-3′) were:
Madreiter-Sokolowski C.T., Gottschalk B., Parichatikanond W., Eroglu E., Klec C., Waldeck-Weiermair M., Malli R, & Graier W.F. (2016). Resveratrol Specifically Kills Cancer Cells by a Devastating Increase in the Ca2+ Coupling Between the Greatly Tethered Endoplasmic Reticulum and Mitochondria. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 39(4), 1404-1420.
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