Psp spin stool kit

Manufactured by Stratec

Sourced in Germany

The PSP Spin Stool Kit is a laboratory equipment designed for sample preparation. It incorporates a compact centrifuge that can be used to separate liquids and sediments in small sample volumes. The kit includes the centrifuge, a selection of rotor buckets, and accessories necessary for its operation.

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Lab products found in correlation

Psp lysis buffer, by Stratec (3 mentions) Glass bead, by Biospec (3 mentions) Zirconia silica bead, by Biospec (3 mentions) Magna lyser instrument, by Roche (2 mentions) Nanodrop nd 1000, by Thermo Fisher Scientific (1 mentions) Magna lyser, by Roche (1 mentions) Magna lyser device, by Roche (1 mentions) Glycerol, by Merck Group (1 mentions) Quant it picogreen dsdna reagent kit, by Thermo Fisher Scientific (1 mentions) Victor3 multilabel counter, by PerkinElmer (1 mentions) Nextera dna flex, by Illumina (1 mentions) Sybr green chemistry, by Roche (1 mentions) Novaseq 6000 sequencing platform, by Illumina (1 mentions) Qubit 2.0 fluorometer, by Thermo Fisher Scientific (1 mentions) Qiaamp dna mini kit, by Qiagen (1 mentions)

Close spelling variants of this product

PSP Spin Stool DNA Plus Kit (54 citations) PSP Spin Stool DNA Kit (35 citations)

7 protocols using psp spin stool kit

Stool DNA Extraction and Purification

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PSP lysis buffer (Stratec Molecular, Berlin, Germany) was added to a sterile vial containing 0.5 g of 0.1 mm zirconia/silica beards and 4 3.0–3.5 mm glass beads (BioSpec, Bartlesville, USA). Frozen stool aliquots were added to the vials. The samples were homogenized in a MagNA Lyser instrument (Roche, Basel, Switzerland) in three cycles of 1 min. at a speed of 5500 rpm. Samples were kept on ice for one minute in between cycles. DNA isolation was continued using the PSP Spin Stool Kit (Stratec Molecular, Berlin, Germany) according to the manufacturers’ instructions. DNA was finally eluted in 200 μl elution buffer. The DNA quantity and quality was measured by NanoDrop ND-1000 (Thermo Scientific, Wilmington, USA).
Samples with a DNA concentration less than 20 ng or an A_260/280 less than 1.8 were subjected to ethanol precipitation to concentrate or further purified, respectively, to meet the quality standards. Ten FecalSwab samples with a total DNA content of less than 400 ng were excluded from this research.

Tedjo D.I., Jonkers D.M., Savelkoul P.H., Masclee A.A., van Best N., Pierik M.J, & Penders J. (2015). The Effect of Sampling and Storage on the Fecal Microbiota Composition in Healthy and Diseased Subjects. PLoS ONE, 10(5), e0126685.

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Fecal Metagenomic DNA Extraction Protocol

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Fecal samples were diluted 10-fold in peptone/water solution (Oxoid, Basingstoke, UK) containing 20% (vol/vol) glycerol (Merck, Darmstadt, Germany) and homogenized by vortexing. They were stored at −20°C until molecular analysis was performed.
For the extraction of metagenomic DNA, 200 μL of diluted feces was added to a 2-mL vial containing 0.5 g of 0.1 mm zirconia/silica beads (BioSpec, Bartlesville, OK, USA), 4 glass beads, 3.0–3.5 mm (BioSpec), and 1.2 mL of lysis buffer from the PSP Spin Stool Kit (Stratec Molecular, Berlin, Germany). Samples were disrupted in a Magna Lyser device (Roche, Basel, Switzerland) in 3 cycles of 1 min. at 5,500 rpm. Subsequently, metagenomic DNA was isolated from the samples by using the PSP Spin Stool Kit according to the manufacturer’s instructions. DNA was eluted in 200 μL elution buffer and stored at −20°C until further analysis.

von Wintersdorff C.J., Penders J., Stobberingh E.E., Lashof A.M., Hoebe C.J., Savelkoul P.H, & Wolffs P.F. (2014). High Rates of Antimicrobial Drug Resistance Gene Acquisition after International Travel, the Netherlands. Emerging Infectious Diseases, 20(4), 649-657.

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Fecal DNA Extraction for Microbiome Analysis

Cited 1 time

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A portion of approximately 200 mg was obtained from frozen fecal samples by cutting on ice to prevent the thawing of samples. Subsequently, DNA was isolated using a combination of bead beating and column-based purification with the PSP spin stool kit (Stratec Molecular, Berlin, Germany) as described previously [39 (link)]. PCR-grade water was used as a negative control. DNA concentrations were determined using a Quant-IT Pico Green dsDNA reagent kit (Invitrogen, New York, NY, USA) using the Victor3 Multilabel Counter (Perkin Elmer, Waltham, MA, USA).

Tedjo D.I., Wilbrink J.A., Boekhorst J., Timmerman H.M., Nienhuijs S.W., Stronkhorst A., Savelkoul P.H., Masclee A.A., Penders J, & Jonkers D.M. (2023). Impact of Sleeve Gastrectomy on Fecal Microbiota in Individuals with Morbid Obesity. Microorganisms, 11(9), 2353.

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Stool DNA Extraction and ITS Amplification

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DNA was extracted from stool samples using a commercially available DNA extraction kit (PSP^Ⓡ Spin Stool Kit, Stratec Molecular, Berlin, Germany) following the manufacturer’s instructions. We amplified a fragment of the ITS region of SSU rRNA genes from the extracted DNA using nested PCR. The amplicons were about 390 base pairs (bps) in length [13 (link)]. The outer primer set was EBITS3 (5′- GGT CAT AGG GAT GAA GAG − 3′) and EBITS4 (5′- TTC GAG TTC TTT CGC GCT C-3′), whereas the inner primer set was EBITS1 (5′- GCT CTG AAT ATC TAT GGC T-3′) and EBITS2.4 (5′- ATC GCC GAC GGA TCA AGT G-3′). The thermal cycling conditions were maintained as follows: initial denaturation at 94 °C for 3 min, 35 cycles of denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s, extension at 72 °C for 40 s, followed by final extension at 72 °C for 10 min. The final PCR products (390 bps) were separated on 2% agarose gel and visualised under a UV transilluminator.

Udonsom R., Prasertbun R., Mahittikorn A., Chiabchalard R., Sutthikornchai C., Palasuwan A, & Popruk S. (2019). Identification of Enterocytozoon bieneusi in goats and cattle in Thailand. BMC Veterinary Research, 15, 308.

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Comprehensive Gut, Oral, and Skin Microbiome Profiling

Cited 1 time

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DNA extraction from the faecal samples is to be obtained by the use of the commercial kit, the PSP Spin Stool Kit (Stratec, USA), with an enzymatic and bead beating step to enhance DNA recovery and concentration. DNA extraction from the oral, vaginal and skin samples are to be obtained using the commercial kit, QIAamp DNA Mini Kit (Qiagen, USA) following previously published methodology.29 (link) DNA concentration is measured using the Qubit 2.0 Fluorometer (Life Technology, USA). Bacterial quantitative PCR analysis of samples will be undertaken to confirm the presence of bacterial DNA, prior to sequence analysis. PCR primers (926F30 and 1062R31 (link)), targeting total bacteria, will be performed using Quantstudio (Thermo Fisher Scientific) using SYBR Green chemistry (Roche). To rule out possible reagent and collection kit contamination, sample collection buffers and double distilled water will be included for DNA extraction, Qubit, qPCR and sequencing. Shotgun metagenomic libraries will be generated with the Illumina Nextera DNA Flex, sequenced on the NovaSeq 6000 Sequencing platform at the UNSW Ramaciotti Centre for Genomics.

Susic D., Davis G., O' Sullivan A.J., McGovern E., Harris K., Roberts L.M., Craig M.E., Mangos G., Hold G.L., El-Omar E.M, & Henry A. (2020). Microbiome Understanding in Maternity Study (MUMS), an Australian prospective longitudinal cohort study of maternal and infant microbiota: study protocol. BMJ Open, 10(9), e040189.

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Fecal DNA Extraction Protocol

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Frozen aliquots of fecal samples were cut on ice to prevent thawing of the fecal samples and approximately 200 mg was added to vials containing PSP lysis buffer (Stratec Molecular, Berlin, Germany), 0.5 g of 0.1 mm zirconia/silica beads and 4 glass beads of 3.0–3.5 mm (BioSpec, Bartlesville, USA). The fecal samples were homogenized in a MagNALyser instrument (Roche, Basel, Switzerland) in three cycles of 1 min at a speed of 5500 rpm. Samples were kept on ice for one minute in between cycles. DNA isolation was continued using the PSP Spin Stool Kit (Stratec Molecular, Berlin, Germany) according to the manufacturers’ instructions. DNA was finally eluted in 200 μl TE-buffer. Negative control samples (PCR grade water) were included in each batch of samples for DNA-isolation, and handled in exactly the same way as the fecal samples, in order to rule out contamination during the isolation procedure.

Tedjo D.I., Smolinska A., Savelkoul P.H., Masclee A.A., van Schooten F.J., Pierik M.J., Penders J, & Jonkers D.M. (2016). The fecal microbiota as a biomarker for disease activity in Crohn’s disease. Scientific Reports, 6, 35216.

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Stool DNA Extraction Protocol

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PSP lysis buffer (Stratec Molecular, Berlin, Germany) was added to a sterile vial containing 0.5 g of 0.1 mm zirconia/silica beads and 4 3.0–3.5 mm glass beads (BioSpec, Bartlesville, USA). 200 mg of frozen stool aliquots were added to the vials. The samples were homogenized in a MagNA Lyser instrument (Roche, Basel, Switzerland) in three cycles of 1 min. at a speed of 5500 rpm. Samples were kept on ice for one minute in between cycles. DNA isolation was continued using the PSP Spin Stool Kit (Stratec Molecular, Berlin, Germany) according to the manufacturers’ instructions. DNA was finally eluted in 200 μl elution buffer.

Mack I., Cuntz U., Grämer C., Niedermaier S., Pohl C., Schwiertz A., Zimmermann K., Zipfel S., Enck P, & Penders J. (2016). Weight gain in anorexia nervosa does not ameliorate the faecal microbiota, branched chain fatty acid profiles, and gastrointestinal complaints. Scientific Reports, 6, 26752.

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