Custom Microbial DNA qPCR Arrays

For real-time PCR-based, customized microbial identification or profiling

Configure at GeneGlobe

Find or custom design the right target-specific assays and panels to research your biological targets of interest.

Custom Microbial DNA qPCR Arrays (Additional Plates)

Cat. No. / ID: 330162

Additional Custom Microbial DNA qPCR Array plates, available at time of initial order

Configure at GeneGlobe to see pricing

Product

Custom Microbial DNA qPCR Arrays (Additional Plates)

Custom Microbial DNA qPCR Arrays

CONFIGURE AT the GeneGlobe

Custom Microbial DNA qPCR Arrays sind für molekularbiologische Anwendungen vorgesehen. Diese Produkte sind nicht zur Diagnose, Prävention oder Behandlung einer Erkrankung vorgesehen.

Configure at GeneGlobe

Find or custom design the right target-specific assays and panels to research your biological targets of interest.

Features

Enables profiling of a customized panel of microbial genes or species
Contains highly sensitive and specific Microbial DNA qPCR Assays
A variety of plate layouts available to suit your research needs
Create your custom plate with our easy-to-use online custom array builder

Product Details

The Custom Microbial DNA qPCR Arrays are ready-to-use 96- and 384-well plates containing user-selected microbial DNA qPCR Assays for detecting microbial species or microbial genes for your particular need. Master Mix is provided with the arrays.

For designing and ordering a Custom Microbial DNA qPCR Array, please go to the custom panel builder in GeneGlobe.

Need a quote for your research project or would you like to discuss your project with our specialist team? Just contact us!

Performance

Linearity and dynamic range

Microbial DNA qPCR Assays display linear amplification across a dynamic range from 10 to 10⁶ copies of DNA template (see figure, ).

Lower limit of quantification (LLOQ)

The LLOQ is the lowest concentration of template that still falls into the linear range of the standard curve (see figure, ). Across all Microbial DNA qPCR Assays, 93% have an LLOQ of <100 gene copies (see figure, ). 92% of microbial identification assays meet this LLOQ, as do 95% of virulence gene assays and 97% of antibiotic resistance gene assays (see figures, , , and ).

Specificity

Each Microbial DNA qPCR Assay is stringently tested to ensure that it detects only one target species or gene (see figure, ). For assays that do detect more than one target, a list of detected targets and in silico predictions is included on the product sheet.
This specificity is maintained even when samples have high species complexity, such as in stool, sputum, and plaque (see figure, ), and is verifiable by sequencing methods (see figure, ).

Reproducibility

Microbial DNA qPCR Assays are highly reproducible, both in intra- and inter-individual variability tests (see figure, ).

See figures

Linearity and sensitivity of Microbial DNA qPCR Arrays.

Limit of detection versus lower limit of quantification.

The lower limit of quantification (LLOQ) for all Microbial DNA qPCR Assays reveals high sensitivity.

The lower limit of quantification (LLOQ) for microbial identification Microbial DNA qPCR Assays reveals high sensitivity.

The lower limit of quantification (LLOQ) for virulence factor gene detection Microbial DNA qPCR Assays reveals high sensitivity.

The lower limit of quantification (LLOQ) for antibiotic resistance gene detection Microbial DNA qPCR Assays reveals high sensitivity.

Microbial DNA qPCR Assays are highly specific.

Microbial DNA qPCR Assays display high sensitivity even in complex metagenomic samples.

Specificity of the Antibiotic Resistance Genes Microbial DNA qPCR Array is verified by pyrosequencing.

Microbial DNA qPCR Arrays generate reliably reproducible results.

Principle

Microbial DNA qPCR Assays are designed to detect bacterial 16S rRNA gene and fungal ribosomal rRNA gene sequences for species identification, as well as virulence factor genes and antibiotic resistance genes using PCR amplification primers and hydrolysis probe-based detection.

Design your own 96- or 384-well microbial DNA assay plate from our selection of predesigned Microbial DNA qPCR Assays. Choose a fully flexible plate layout or select from several convenient predefined layouts. Use our easy-to-use online plate configurator to position the predesigned Microbial DNA qPCR Assays of interest into your plate. The arrays will be delivered ready-to-use, with one single assay per well, sufficient for a 20 µl reaction volume. The Master Mix is provided with the arrays.

The table below displays the many different plate layout combinations available for each plate format. Select the plate layout that suits your needs; for example, configure a 96-well plate with 48 assays to be used with 2 samples, or a 96-well plate with 12 assays to be tested with 8 samples.

We highly recommend that you include controls on each Custom Microbial DNA qPCR Array. Controls are available for testing for the presence of fungal DNA, bacterial DNA and host genomic DNA and for assessing the success of the PCR reaction, so you can be confident in the results.

Plate format	Assays x samples per plate	Plate pack size (minimum)
96-well plate or Rotor-Disc 100	8 assays x 12 samples	12 plates
	12 assays x 8 samples
	16 assays x 6 samples
	32 assays x 3 samples	24 plates
	48 assays x 2 samples
	96 assays x 1 samples
384-well plate	8 assays x 48 samples	6 plates
	12 assays x 32 samples
	16 assays x 24 samples
	24 assays x 16 samples	6 plates
	32 assays x 12 samples
	48 assays x 8 samples
	64 assays x 6 samples
	96 assays x 4 samples
	128 assays x 3 samples
	192 assays x 2 samples	24 plates
	384 assays x 1 samples	24 plates

Procedure

The Microbial DNA qPCR Array procedure is simple and can be carried out in any laboratory with a real-time PCR instrument. DNA is isolated using the QIAamp kit that is suited for the sample type, and then mixed with the appropriate Microbial qPCR Mastermix. The mixture is aliquotted across the array plate, and real-time PCR is performed to obtain the raw C_T values for each assay. The complimentary data analysis software is then used to profile genes or species in the sample.

Applications

Custom Microbial DNA qPCR Arrays are highly suited for the rapid and accurate profiling of a custom panel of microbial species, virulence factor genes, or antibiotic resistance genes. For example, a microbial species identification panel assembled by our experts was used to explore the underlying causes of bacterial vaginosis (see figures, and ), while a panel for antibiotic resistance genes identified these genes in samples from the gut or from sewage (see figures, and ).

See figures

The Vaginal Flora Microbial DNA qPCR Array provides accurate profiling for cervical swab samples.

Vaginal samples positive for Gardnerella vaginalis also show changes in commensal and bacterial vaginosis-related microbes compared to healthy samples.

The Microbial DNA qPCR Array screens gut microbiota for the presence of antibiotic resistance genes.

The Antibiotic Resistance Genes Microbial qPCR Array identified antibiotic resistance genes in sewage samples.

Supporting data and figures

Linearity and sensitivity of Microbial DNA qPCR Arrays.

Linearity and sensitivity for each Microbial DNA qPCR Array was determined using synthetic templates over a 6 log serial dilution ranging from 1 copy to 1 million copies. The following are representative results for all the qPCR assays. [A] shows the real-time amplification curves of the KPC antibiotic resistance gene qPCR assay. In [B], a standard curve was prepared that shows that the primer efficiency equals 103% (calculated from slope = –3.3236) and the correlation coefficient is 0.9983, indicating optimum performance for the KPC qPCR assay. All Microbial DNA qPCR Assays have primer efficiencies between 80–120% and correlation coefficients (R)>0.995.