Mycycler real time pcr system

Manufactured by Bio-Rad

Sourced in Denmark

The MyCycler real-time PCR system is a laboratory instrument designed for the amplification and detection of DNA and RNA sequences. It utilizes the real-time PCR technique to monitor the progress of a PCR reaction in real-time. The system is capable of performing quantitative and qualitative analysis of nucleic acid samples.

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Lab products found in correlation

Iscript advanced cdna synthesis kit, by Bio-Rad (1 mentions) Rneasy mini kit, by Bio-Rad (1 mentions) Iscript cdna synthesis kit, by Bio-Rad (1 mentions) Iq sybr green supermix, by Bio-Rad (1 mentions) Nanodrop, by Thermo Fisher Scientific (1 mentions) Aurum total rna mini kit, by Bio-Rad (1 mentions)

Close spelling variants of this product

MyCycler (172 citations) Real-Time System (61 citations) MyCycler PCR system (7 citations) MyCycler PCR (2 citations) MyCycler system (2 citations)

2 protocols using mycycler real time pcr system

Osteogenic Differentiation of hASCs

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hASCs were seeded on a 12-well plate at a density of 2×10⁵ cells/well and incubated overnight. Cells were cultured for 14 days in osteogenic differentiation medium including test solution, and the medium was exchanged twice with a fresh one after 5 and 10 days. Then, total RNA was isolated with an RNeasy minikit (Bio-Rad) according to the manufacturer’s protocol. Then, cDNA was synthesized from extracted RNA using an iScript advanced cDNA-synthesis kit (Bio-Rad). All primers used were produced by DNA Technology (Aarhus, Denmark). The total volume of each sample was 20 μL, and samples were analyzed in duplicate with a MyCycler real-time PCR system (Bio-Rad). The thermocycling program consisted of an initial step of 3 minutes at 95°C and 40 cycles of 15 seconds at 95°C and an annealing/extension temperature of 60°C for 30 seconds. Sequences of the primer sets were:
Osteocalcin (OCN) forward 5ʹ-GAG CCC CAG TCC CCT ACC C-3ʹ, reverse 5ʹ-GCC TCC TGA AAG CCG ATG TG-3ʹ
Osteopontin (OPN) forward 5ʹ-TGA TGG CCG AGG TGA TAG TGT GGT-3ʹ, reverse 5ʹ-CCT GGG CAA CGG GGA TGG-3ʹ
Peptidylprolyl isomerase A (PPIA) forward 5ʹ-TCC TGG CAT CTT GTC CAT G-3ʹ, reverse 5ʹ-CCA TCC AAC CAC TCA GTC TTG-3ʹ.
Fold-change gene-expression levels were recorded using the ΔΔCt method, in which the data were normalized with the expression level of PPIA.

Jahed V., Vasheghani-Farahani E., Bagheri F., Zarrabi A., Fink T, & Lambertsen Larsen K. (2019). Enhanced Cellular Uptake Of Phenamil Through Inclusion Complex With Histidine Functionalized β-Cyclodextrin As Penetrative Osteoinductive Agent. International Journal of Nanomedicine, 14, 8221-8234.

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Quantitative Real-Time PCR Assay

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Total cellular RNA was extracted from cells at indicated time points using an Aurum total RNA mini kit (Bio-Rad, Copenhagen, Denmark) according to the manufacturer’s instructions. The purity and concentration were determined on the basis of the 260/280 optical density (OD) ratio using spectrophotometry (NanoDrop; Thermo Fisher Scientific, Wilmington, DE, USA). Complementary DNA (cDNA) was synthesized using iScript cDNA synthesis kit (Bio-Rad, Copenhagen, Denmark). All primer sequences were designed using software Primer3 and were produced by DNA Technology (Aarhus, Denmark). PCR reactions were monitored and performed on a My-Cycler real-time PCR system (Bio-Rad, Copenhagen, Denmark). IQ SYBR Green Supermix (Bio-Rad, Copenhagen, Denmark) was used in each reaction. Reactions were performed for 40 amplification cycles as follows: 3 min at 95 °C, 10 s at 95 °C for DNA denaturation and 30 s for annealing and extension at respective annealing temperature (Table 1). The expression level for each gene of interest was normalized to that of the β-actin. The relative ratio of each gene was obtained by dividing the mRNA expression at week 2 by that of week 0. A blank without cDNA was included as a negative control.

Wang F., Zachar V., Pennisi C.P., Fink T., Maeda Y, & Emmersen J. (2018). Hypoxia Enhances Differentiation of Adipose Tissue-Derived Stem Cells toward the Smooth Muscle Phenotype. International Journal of Molecular Sciences, 19(2), 517.

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