Mastercycler 5330

Manufactured by Eppendorf

Sourced in Germany

The Mastercycler 5330 is a thermal cycler designed for PCR (Polymerase Chain Reaction) amplification of DNA samples. It features a compact, robust design and provides precise temperature control for efficient DNA amplification.

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Lab products found in correlation

Cdna synthesis kit, by Thermo Fisher Scientific (1 mentions) Tripure reagent, by Roche (1 mentions) Taq dna polymerase, by Roche (1 mentions) Qiamp dna mini kit, by Qiagen (1 mentions) Sybr green, by Thermo Fisher Scientific (1 mentions) Pcr grade water, by Thermo Fisher Scientific (1 mentions)

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Mastercycler (595 citations)

4 protocols using mastercycler 5330

RNA Extraction and Gene Expression Analysis

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Total RNA was extracted from treated or co-cultured hWJ-MSC derived cells using TRIpure reagent (Roche, Germany) according to manufacturer instructions. Afterwards, cDNA was generated using 1 μg of the isolated total RNA by a cDNA synthesis kit (Invitrogen, USA) according to the kit instructions.
Appropriated primer sets for PGC and spermatogonial specific markers, which are listed in Table 1, including POU5F1, Fragilis, DDX4, Plzf and Piwil2 (Mili), Stra8, Dazl, β1- and α6-integrins (ITΒ1, ITA6) were designed using Primer3 software. PCR reactions were performed using Taq DNA polymerase (Roche, Germany). The PCR mixture contained 1 µl template cDNA, 0.4 µM of each primer (1 µl), 0.2 mM of dNTPs (0.5 µl), 0.625 unit/25 µl reaction of Taq DNA polymerase (0.125 µl), 1.5 mM MgCl₂ (0.75 µl) and 1X PCR Buffer (2.5 µl) in a total volume of 25 µl with distilled water. In an Eppendorf thermal cycler (Mastercycler® 5330) PCR reactions were performed as follows: 94°C for 3 min, 35 cycles of PCR at 94°C for 30 sec, 60 °C for 30 sec, 72 °C for 1 min, and 72 °C for 5 min. GAPDH was used as a control housekeeping gene.

nejad N.A., Amidi F., Hoseini M.A., Nia K.N., Habibi M., Kajbafzadeh A.M., Mazaheri Z, & Yamini N. (2015). Male germ-like cell differentiation potential of human umbilical cord Wharton’s jelly-derived mesenchymal stem cells in co-culture with human placenta cells in presence of BMP4 and retinoic acid. Iranian Journal of Basic Medical Sciences, 18(4), 325-333.

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Multiplex PCR for STEC and E. coli O157:H7

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Isolates from 65 cheese samples were tested for the presence of STEC and E. coli O157:H7 strains by determining STX (VT1 and VT2) and eae genes in DNA using known primers (Table-2) in multiplex polymerase chain reaction (PCR) according to the methodology described in the Terrestrial Manual of the World Organization for Animal Health [23 ].
For DNA extraction from E. coli colonies, the QIAmp DNA Mini Kit (Qiagen, Germany) was used according to the manufacturer’s instructions. DNA concentration was measured using a NanoDrop 1000 spectrophotometer at a wavelength of 260 nm, and a qualitative assessment of DNA was performed by electrophoresis.
The multiplex PCR reaction was performed in a total volume of 48 µL, including 3 mM MgCl₂, 3 mM buffer KCL, 3 mM dNTP 2 mM, 25 pmol of each primer (forward and reverse primers for VT1, VT2, and eaeA), 0.2 Taq polymerase, and 3 mL (40–260 ng/mL) DNA. The amplification reaction was carried out using a DNA thermal cycler (Eppendorf Mastercycler 5330), which included denaturation for 2 min at 94°C; 28 cycles: 94°C–1 min, 62°C–1.5 min, and 72°C–2 min; and final elongation at 72°C–5 min. Amplified samples were electrophoresed in 1.5% agarose gel and stained with ethidium bromide. We used a molecular weight marker with a step length of 100 bp as the size standard.

Kuzeubayeva A., Ussenbayev A., Aydin A., Akanova Z., Rychshanova R., Abdullina E., Seitkamzina D., Sakharia L, & Ruzmatov S. (2024). Contamination of Kazakhstan cheeses originating from Escherichia coli and its resistance to antimicrobial drugs. Veterinary World, 17(2), 361-370.

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Detecting Toxoplasma gondii B1 Gene

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DNA thermo-cycler (Eppendorf Master cycler 5330, Hamburg, Germany) was applied in PCR reactions. T. gondii B1 gene was perceived in DNA samples by means of the method conveyed formerly [19] . Table 1 signifies the PCR circumstances applied for detection of T. gondii B1 gene. Visualization was performed by means of electrophoresis on agarose gel (2%in 1× TBE buffer stained with SYBR Green (Thermo Fisher Scientific, Germany). Negative control (PCR grade water (Thermo Fisher Scientific, Germany) and positive control (positive DNA for the B1 gene obtained from the Faculty of Veterinary Medicine, University of Tehran) were applied in PCR reactions.

Amroabadi M.A., Rahimi E., & Shakerian A. (2020). Seasonal and Age Distribution of Toxoplasma gondii in Milk of Naturally Infected Animal Species and Dairy Samples. Egyptian Journal of Veterinary Science, 51(2), 171-180.

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Bacterial Gene Amplification Using PCR

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The PCR was performed using 2 µL of the bacterial lysates and primer pairs to amplify the target genes individually. PCR was performed in 20-µL reactions. Each reaction contained 1 pmol/µL of each forward and reverse primer, 160 uM dNTPs, 2.5 mM MgCl 2 , 1 × PCR buffer, 0.75 U Taq DNA polymerase and 10 ng DNA template. PCR was performed using the Mastercycler 5330 (Eppendorf, Hamburg, Germany), with an initial denaturation at 94 o C for 3 min; 30 cycles of denaturation at 94 o C for 30 s, annealing for 30 s at the TA of the primer pairs, and extension at 72 o C for 30 s; followed by an extra cycle of annealing at the TA for 30 s and a final extension at 72 o C for 5 min. The PCR products were electrophoresed through 2% low EEO agarose gel (Promega, Madison, USA) stained with ethidium bromide, at 100 volts for 60 min. PCR products were seen using an image analyzer.

Al-Talib H., Zuraina N., Kamarudin B, & Yean C.Y. (2015). Genotypic variations of virulent genes in Enterococcus faecium and Enterococcus faecalis isolated from three hospitals in Malaysia. Advances in clinical and experimental medicine : official organ Wroclaw Medical University, 24(1).

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