Assays for quantifying prokaryotes

£85 per single assay

To order please send a Purchase Order to finance@qstandard.co.uk

 

Enterobacter_cloacae_01

 

Bacteria

This assay is designed to amplify the single copy bacterial gene rpoB. does not amplify DNA from archaea. This assay uses a number of degenerate primers designed against a conserved region of the gene to amplify bacterial rpoB. It has been tested in several individual bacterial cultures including Staphylococcus albus, Micrococcus luteus, Escherichia coli, Bacillus subtilis, Bacillus megaterium and Bacillus mycoides, as well as in mixed baterial populations such as oral biofilms and environmental samples. When used in a mixed sample, amplicons generated may differ in sequence but not in product size. This does not affect the accuracy of quantification.

The stock assay, 2_rpoB_G01, contains a standard derived from several species for use when quantifying samples of unknown bacterial composition, but standards for individual species can be provided if preferred, please email info@qstandard.co.uk.

Gene symbol Kingdom Product code Amplicon length (bp) Amplicon %GC
rpoB bacteria 2_rpoB_G01 181 varies

 

    ? Primers have been designed to avoid secondary structure (self-annealing)
    ? No secondary structure in the amplicon for any assay (positive delta G)
    ? Specificity has been checked by melt peak analysis and the presence of only a single band on a gel
    ? Standard curves down to 10 copies per reaction have been run for each assay and all show excellent linearity (R² >0.99) Efficiency (>95%) and sensitivity (repeat of 10 copies consistently detected)
    ? Primers have been tested at an annealing temperature of 57°C. Higher annealing temperatures (up to 60°C) are possible, but require individual validation

 

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