MBD6000

5R-Plex kit

Ultra-sensitive 16S NGS assay for degraded and low biomass DNA

• UNSPSC-Code: 41106302
• NACRES: NA.84

Eigenschaften

• Qualitätsniveau: 200
• Verwendung: Preparing 96 samples for sequencing
• Methode(n): DNA amplification: suitable; DNA sequencing: suitable
• Versandbedingung: dry ice
• Lagertemp.: -10 to -25°C

Beschreibung

Allgemeine Beschreibung

The 16S ribosomal RNA gene (16S rRNA) is a common bacterial marker, used as a target for microbial community profiling in microbiome metagenomic studies. The gene is composed of nine variable regions (V1-V9) interspersed between conserved regions. The most common 16S rRNA NGS assays target one or two regions using a single set of primers (e.g., V3-V4). However, this approach usually results in a poor bacterial detection and classification rate when the DNA input is of low-quality or when applying it on extremely low biomass samples.  The 5R-PLEX kit offers a complete solution to challenging biological samples where the standard 16S sequencing protocols fail to perform.   Sample types include:

• Formalin-Fixed, Paraffin-Embedded (FFPE) tissue[1]
• Cancerous tumor tissue 
• Degraded or damaged DNA  
• Low biomass samples 
• Fossil-derived and ancient DNA

The 5R-PLEX kit was developed in collaboration with Dr. Ravid Straussman of the Weissmann Institute, based off a technique that was featured in his Science publication focused on the human tumor microbiome characterization (Science2020 May 29;368(6494):973-980. doi: 10.1126/science.aay9189) . The kit targets five short variable regions along the 16S rRNA gene (V2, V3, V5, V6, V8) that are co-amplified in a multiplexed PCR within a single tube.   After sequencing, the output data is uploaded to our M-CAMPTM platform (Microbiome Computational Analysis for Multi-omics Profiling) and analyzed using the unique 5R-PLEX algorithm[2]. This new analysis approach computationally combines the amplified regions to reconstruct one coherent high-resolution microbial profile. Read more about our kit here.

Leistungsmerkmale und Vorteile

• The 5R-PLEX Kit provides high profile sequencing from degraded, damaged or low biomass samples
• Multiplexed NGS assay targets 5 variable regions of the 16S rRNA gene in a single primer pool
• Includes positive control for trouble shooting experiments.
• The kit can detect as low as 1pg of highly degraded bacterial DNA with an average fragment size of 260bp.
• Kit includes low bioburden reagents
• Includes exclusive access to M-CAMP^™ platform featuring an innovative algorithm designed to reconstruct a single coherent microbial profiling and perform comprehensive analysis.

Angaben zur Herstellung

The shelf life of all reagents provided in the kit is 12 months when stored properly. Store all components at -20 °C.

Sonstige Hinweise

• 5R-PLEX PCR1 Primer mix- 25 μL
• 5R-PLEX PCR2 Forward Primer mix- 25 μL
• 5R-PLEX index plate 96-plate
• Water, microbial DNA-free 10X- 1.5 mL
• HF DNA Polymerase- 0.1 mL
• 5X HF buffer 2 mL
• dNTP′s- 0.2 mL
• Elution Buffer (EB), microbial DNA-free- 8 mL
• 5R-PLEX Positive Control (10 ng/μL)- 30 μL

Kit component details can be found in the User guide.

Rechtliche Hinweise

M-Camp is a trademark of Merck KGaA, Darmstadt, Germany

Sicherheitshinweise

• H-Sätze: H412
• P-Sätze: P273 - P501
• Gefahreneinstufungen: Aquatic Chronic 3
• Lagerklassenschlüssel: 10 - Combustible liquids