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Magna pure bacteria lysis buffer

Manufactured by Roche
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The MagNA Pure Bacteria Lysis Buffer is a lab equipment product designed to lyse bacterial cells, releasing their cellular contents for further processing. The buffer is a key component in nucleic acid extraction and purification workflows.

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Lab products found in correlation

Magna pure compact nucleic acid isolation kit 1, by Roche (6 mentions) Magna pure compact nucleic acid isolation kit, by Roche (5 mentions) Lightcycler 2.0 instrument, by Roche (3 mentions) Fast start light cycler 480 hybprobes master kit, by Roche (3 mentions) Magna pure compact, by Roche (3 mentions) Magna pure compact instrument, by Roche (3 mentions) Magna lyser instrument, by Roche (2 mentions) Miseq benchtop sequencer, by Illumina (2 mentions) Nextera xt dna library preparation kit, by Illumina (2 mentions) One step real time kit, by Thermo Fisher Scientific (1 mentions) Lyticase, by Merck Group (1 mentions) Magna lyser green beads, by Roche (1 mentions) Magna pure dna isolation kit 3, by Roche (1 mentions) Magna pure lc instrument, by Roche (1 mentions) Magna lyser, by Roche (1 mentions) Septifast lys kit, by Roche (1 mentions) Magna pure compact system, by Roche (1 mentions) Bactec fx, by BD (1 mentions) Proteinase k, by Merck Group (1 mentions) Proteinase k, by Roche (1 mentions)

Spelling variants of this product

Lysis buffer (81 citations) MagNA Pure (79 citations) Magna Pure Lysis Buffer (12 citations)

15 protocols using magna pure bacteria lysis buffer

1

SARS-CoV-2 RNA Detection in Tissue Samples

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After the retrieval, tissues were transferred in antibiotic solution made up with Gentamicin, Vancomycin and Meropenem and maintained at + 4 °C following the validated FBTV internal procedures (Serafini et al. 2016 (link); Montagner et al. 2018 (link); Paolin et al. 2018 (link)).
Before the analysis, tissues were kept at + 4 °C.
The tissues were manually dissected into few sections of approximately 2 mm and treated over night with MagNA Pure Bacteria Lysis Buffer (Roche). Detection of SARS-CoV-2 RNA was performed by an in-house real-time RT–PCR method, which was developed according the protocol and the primers and probes designed by CDC that targeted the gene encoding nucleocapsid (N2) of SARS-CoV-2 and RNA-dependent RNA polymerase. Real-time RT–PCR assays were performed in a final volume of 25 μl, containing 5 μl of purified nucleic acids, using One Step Real Time kit (Thermo Fisher Scientific) and run on ABI 7900HT Fast Sequence Detection Systems (Thermo Fisher Scientific).
Montagner G., De Vettor R., Favaretto F., Vici D., Del Vecchio C., Franchin E., Trojan D, & Feltrin G. (2022). Impact of Sars-CoV-2 pandemic on the Veneto Region multitissue bank activity. Cell and Tissue Banking, 23(4), 825-832.
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2

Stability Assessment of STR Markers in Candida

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Isolates were grown on Sabouraud agar plates at 35°C. To test the stability of STR markers, 20 colonies of isolates 10-08-01-01 and VPCI 247/P/15 were clonally expanded for 5 generations on Sabouraud agar plates. For DNA sequencing, strains were resuspended in a vial with 400 μl MagNA Pure Bacteria lysis buffer and MagNA Lyser green beads and mechanically lysed for 30 s at 6,500 rpm using the MagNA Lyser (all Roche Diagnostics GmbH, Mannheim, Germany). Subsequently, DNA was extracted and purified with the MagNA Pure LC instrument and the MagNA Pure DNA isolation kit III (Roche Diagnostics), according to the recommendations of the manufacturer. For STR analysis, strains were resuspended in 50 μl physiological salt, and after the addition of 200 U of lyticase (Sigma-Aldrich, St. Louis, MO, USA) and incubation for 5 min at 37°C, 450 μl physiological salt was added. The sample was then incubated for 15 min at 100°C and cooled down to room temperature.
de Groot T., Puts Y., Berrio I., Chowdhary A, & Meis J.F. (2020). Development of Candida auris Short Tandem Repeat Typing and Its Application to a Global Collection of Isolates. mBio, 11(1), e02971-19.
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3

Bacterial DNA Extraction and Identification

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DNA Extraction: DNA was extracted from overnight colonies of a bacterial culture grown on Cetrimide agar. This was resuspended in Roche MagNA Pure Bacteria Lysis Buffer, vortexed briefly, heated at 95°C for 10 minutes, and pelleted by centrifugation at 13000 g for 10 minutes. Four hundred microliters were used as a specimen in the MagNA Pure Compact (MPC) System (Roche Applied Science, Indianapolis), using MPC Nucleic Acid isolation kit 1 according to the manufacturer's instructions. Elution tubes containing 200 µl purified nucleic acids were stored at ‒80°C until further use. The LightCycler 2.0 instrument (Roche Applied Science, Germany) and Fast start LightCycler 480 HybProbes Master Kit (Roche Diagnostics, USA) were used for polymerase reaction. Specific primers and probes (Table 1) designed by TIB Molbiol (Germany) targeting the gene, species-specific gyrB, were amplified by singleplex real-time polymerase chain reaction (rPCR) following the protocol shown in Table 2.
Hosu M.C., Vasaikar S., Okuthe G.E, & Apalata T. (2021). Molecular Detection of Antibiotic-Resistant Genes in Pseudomonas aeruginosa from Nonclinical Environment: Public Health Implications in Mthatha, Eastern Cape Province, South Africa. International Journal of Microbiology, 2021, 8861074.
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4

Biofilm Extraction and DNA Isolation

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Enamel-dentin slabs (T0 and T3) were glued into the lids of 1.5 ml Eppendorf tubes with epoxy resin adhesive that covered all sides except the standardized surface with the biofilm on it. Sterile and DNA-free glass beads and 200 μl ultra-pure water were inserted, and the biofilm in the vials was shredded for 2 min. For total DNA isolation, the lysate was mixed with 380 μl of MagNA Pure Bacteria Lysis Buffer (Roche Applied Science, Mannheim, Germany) together with 20 μl of proteinase K solution (20 mg/ml) and incubated at 65°C for 10 min. Proteinase K was heat-inactivated at 95°C for another 10 min. The liquid samples were transferred to MagNA Pure Compact Sample Tubes. DNA isolation was performed on the MagNA Pure Compact instrument according to manufacturer instructions using the MagNA Pure Compact Nucleic Acid Isolation Kit I and following the bacteria purification protocol (Roche Diagnostics, Mannheim, Germany). The DNA was eluted in 50 μl elution buffer and stored at −20°C pending further processing.
Klug B., Santigli E., Westendorf C., Tangl S., Wimmer G, & Grube M. (2016). From Mouth to Model: Combining in vivo and in vitro Oral Biofilm Growth. Frontiers in Microbiology, 7, 1448.
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5

Automated Bacterial DNA Extraction Protocol

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Prior to DNA extraction, bacterial suspensions were pretreated by mixing 180 μl of MagNA Pure Bacteria Lysis Buffer (Roche Applied Science, Penzberg, Germany), 20 μl of proteinase K solution (20 g/l), and 200 μl of the bacterial suspension. The mixture was incubated at 65 °C for 10 min and at 95 °C for another 10 min. After transferring 400 μl of the suspension to the MagNA Pure Compact sample tube, the DNA was extracted through a fully automated procedure using the MagNA Pure Compact instrument (Roche) with the MagNA Pure Compact Nucleic Acid Isolation Kit I (Roche). The DNA bacterium purification protocol was chosen with an elution volume of 50 μl. The extraction was performed in duplicate, and eluates were pooled to a final volume of 100 μl. These comparable eluates were stored at ms 70° C pending analysis.
Santigli E., Leitner E., Wimmer G., Kessler H.H., Feierl G., Grube M., Eberhard K, & Klug B. (2016). Accuracy of commercial kits and published primer pairs for the detection of periodontopathogens. Clinical Oral Investigations, 20(9), 2515-2528.
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6

Bacterial Genomic DNA Extraction

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Genomic DNA for Sanger sequencing of the partial 16S rRNA gene and Illumina whole-genome sequencing was extracted from overnight bacterial colonies. Mechanical lysis of bacterial cells was performed using the Septifast Lys kit (Roche Diagnostics, Mannheim, Germany), MagNa Pure bacteria lysis buffer (Roche), and the MagNA Lyser instrument (Roche), followed by extraction on a MagNaPure Compact instrument (Roche) using the MagNA Pure Compact nucleic acid isolation kit I (Roche) according to the manufacturer’s instructions.
Sivertsen A., Dyrhovden R., Tellevik M.G., Bruvold T.S., Nybakken E., Skutlaberg D.H., Skarstein I, & Kommedal Ø. (2022). Escherichia marmotae—a Human Pathogen Easily Misidentified as Escherichia coli. Microbiology Spectrum, 10(2), e02035-21.
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7

Fungal DNA Extraction using MagNA Lyser

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DNA was extracted from A. fumigatus cultures a minimum of 4 days old. A conidial suspension was created in MagNA® Pure Bacteria Lysis Buffer (Roche) and transferred to a MagNa® Lyser Green Beads tube (Roche) for homogenization with the MagNa® Lyser Instrument (Roche). We then extracted 400 μL of supernatant using the MagNa® Pure Compact Nucleic Acid Isolation Kit and a MagNa® Pure Extraction Instrument (Roche Diagnostics), according to the manufacturer’s instructions.
Guegan H., Chevrier S., Belleguic C., Deneuville E., Robert-Gangneux F, & Gangneux J.P. (2018). Performance of Molecular Approaches for Aspergillus Detection and Azole Resistance Surveillance in Cystic Fibrosis. Frontiers in Microbiology, 9, 531.
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8

DNA Extraction Using Roche MagNA Pure

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DNA extraction was done using Roche MagNA Pure Bacteria Lysis Buffer, MagNA Pure Compact Nucleic Acid Isolation Kit 1 in MagNA Pure Compact System (Roche Applied Science, Indianapolis).
Vasaikar S., Obi L., Morobe I, & Bisi-Johnson M. (2017). Molecular Characteristics and Antibiotic Resistance Profiles of Klebsiella Isolates in Mthatha, Eastern Cape Province, South Africa. International Journal of Microbiology, 2017, 8486742.
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9

Automated Bacterial DNA Extraction

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Bacterial DNA was prepared by first placing the paper points in a mixture of 380 μl of MagNA Pure Bacteria Lysis Buffer (Roche Applied Science, Mannheim, Germany) and 20 μl of proteinase K solution (20 g/l). The suspension (including the paper points) was incubated at 65°C for 10 min and subsequently at 95°C for another 10 min. After removal of the paper points, the suspension was transferred into the MagNA Pure Compact Sample Tube (Roche Diagnostics, Mannheim, Germany). Automated DNA extraction was performed on the MagNA Pure Compact instrument (Roche) according to manufacturer instruction using the MagNA Pure Compact Nucleic Acid Isolation Kit I (Roche Diagnostics, Mannheim, Germany). Prior to the start of DNA extraction, the instrument adds the heterologous IC automatically. For extraction of bacterial DNA, the DNA Bacteria Purification protocol was used according to manufacturer instructions. DNA was eluted in 50 μl dH2O and stored at −20°C until use.
Santigli E., Trajanoski S., Eberhard K, & Klug B. (2017). Sampling Modification Effects in the Subgingival Microbiome Profile of Healthy Children. Frontiers in Microbiology, 7, 2142.
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10

Rapid Multiplex PCR for Carbapenemase Detection

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Twenty-three blood cultures spiked with genotypically characterized isolates were tested, plus four blood cultures from bacteraemic ill patients. Spiked blood cultures were generated by inoculating blood culture flasks (Bactec TM, Aerobic/F Culture Vials, Becton Dickinson, Germany) containing 10 mL of human blood and 200 μL of a suspension of the respective isolate derived from a fresh overnight culture (with a turbidity equivalent to that of a 0.5 McFarland standard). The spiked isolates were four blaKPC, three blaVIM, four blaNDM, four blaIMP, five blaOXA-48 and three negative controls: one blaSHV-12, one blaCMY-2 and one blaCTX-M-15. Incubation was performed in an automated system (Bactec FX, Becton Dickinson, Germany) until the flask was flagged positive. In blood cultures obtained directly from the patients, the procedure was applied after the flask flagged was positive and once the GeneXpert procedure had been positive.
To 200 μL of sample (PBS from swab or blood culture) were added 180 μL of MagNa Pure Bacteria Lysis Buffer (Roche Diagnostics) and 20 μL of Proteinase K (20 μg/μL, Sigma–Aldrich, Germany). After incubation at 65°C for 10 min the solution was submitted to automatic DNA extraction, using the MagNa Pure Compact Nucleic Acid Isolation Kit (Roche Diagnostics). After 25 min, the extraction was complete and the multiplex PCR was performed as explained above.
Oviaño M., Torres I., González M, & Bou G. (2016). Evaluation of a novel procedure for rapid detection of carbapenemase-producing Enterobacteriaceae (CPE) using the LightMix® modular carbapenemase kits. Journal of Antimicrobial Chemotherapy, 71(12), 3420-3423.
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