Fastprep precellys24

Manufactured by Bertin Technologies

The FastPrep Precellys24 is a high-performance sample preparation instrument designed for efficient homogenization and disruption of a wide range of biological samples, including tissues, cells, and microorganisms. It utilizes a rapid, high-speed shaking motion to break down samples, preparing them for subsequent analysis.

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Lab products found in correlation

Dna engine peltier thermal cycler, by Bio-Rad (2 mentions) Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions) High prime, by Roche (1 mentions) Sybr premix ex taq tli rnaseh plus green with rox, by Takara Bio (1 mentions) Recombinant rnasin, by Promega (1 mentions) Turbo dnase, by Thermo Fisher Scientific (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Improm 2 reverse transcriptase, by Promega (1 mentions) Maxima reverse transcriptase, by Thermo Fisher Scientific (1 mentions) Dnase, by Roche (1 mentions) Rneasy kit, by Qiagen (1 mentions)

2 protocols using fastprep precellys24

RNA Extraction and qRT-PCR Analysis

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RNA was purified from exponentially growing cells using RNeasy®Mini Kit (Qiagen) following the manufacturer instructions. Previously, yeast cells were broken in a FastPrep Precellys24 (Bertin technologies) with glass beads in the recommended kit buffer. The sample was incubated with Turbo DNase (Ambion) and after DNase inactivation and incubation with oligo dT, cDNA was obtained with Improm-II® Reverse Transcriptase and Recombinant RNasin® (Promega) following the manufacturer instructions. The cDNA was analized by quantitative RT-PCR in a DNA Engine Peltier Thermal Cycler (Bio Rad) using the SYBR Premix Ex Taq Tli RNase H Plus Green with ROX (Takara).
Northern analysis were carried out as previously described [19]. CLN2 and ACT1 mRNA were detected using ³²P-labeled probes obtained with HighPrime (Roche) according to the manufacturer instructions.

Quilis I., Taberner F.J., Martínez-Garay C.A., Alepuz P, & Igual J.C. (2019). Karyopherin Msn5 is involved in a novel mechanism controlling the cellular level of cell cycle regulators Cln2 and Swi5. Cell Cycle, 18(5), 580-595.

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Quantitative RT-PCR Analysis of Yeast mRNA

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Approximately 2 × 10⁸ cells were broken in a FastPrep Precellys24 (Bertin technologies). Total RNA was extracted using Qiagen RNeasy kit according to the manufacturer’s protocol. RNA was quantified with NanoDrop 2000 Spectrophotometer (Thermo Scientific). 10 μg of RNA were incubated with DNase (Roche) and after DNase inactivation and incubation with oligo dT, cDNA was obtained with reverse transcriptase Maxima Reverse Transcriptase (Thermo Scientific). cDNA was analyzed in triplicate by quantitative RT-PCR with DNA Engine Peltier Thermal Cycler (BioRad) using the NZYSpeedy qPCR Green Master Mix. mRNA levels of genes of interest were quantified relative to ACT1 mRNA by the ΔCt method.

Ros-Carrero C., Spiridon-Bodi M., Igual J.C, & Gomar-Alba M. (2024). The CDK Pho85 inhibits Whi7 Start repressor to promote cell cycle entry in budding yeast. EMBO Reports, 25(2), 745-769.

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