Since this kit is so sensitive, avoiding possible sources of contamination is key for a successful qPCR.
– Work in a clean environment and decontaminate the benchtop and pipettes with a 10% bleach solution.
– Wipe down the areas with ethanol to remove the trace amounts of bleach and allow the area to try.
– Use filtered tips.
– Take care to ensure that pipetting is efficient and accurate. Calibrate your pipettes regularly.
– Add standards directly into the premix and pipet up and down to flush out any remaining liquid from the tip.
– Avoid pipetting onto the walls as this may skew the results.
– The SYTO9 dye present within the premix is sensitive to light, so working quickly and diligently is recommended.

Our qPCR Premixes does not contain ROX Reference Dye. Ensure that the ROX channel of your device is switched off.

Our qPCR Premixes contains Syto9. You can use the SYBR Green channel for signal detection.

No, kits are not certified.

-Bacterial Standards are generated from E. coli strain JM109.
-Standards are QC tested serial dilutions of genomic DNA.

E. coli strain JM109 is a derivative of strain K12 and is also called K12 JM109.
The genome size of the original genome of H12 is 4.64 Mb (ref).
Strain JM109 genome is 5.5 Mb and contains 7 copies of 16S rRNA gene.
Ref: https://www.ncbi.nlm.nih.gov/pubmed/9278503

The standards included in the Femto Bacterial DNA Quantification kit are serial dilutions of E. coli genomic DNA. Each E. coli genome has about 7 copies of 16S rRNA gene. One can back calculate from the amount of genomic DNA to determine genomic copies, and then multiply by a factor of 7 to determine total copies of the 16S rRNA gene.

Genome copy # = DNA (g) / (g_to_bp const. x genome size)
With DNA (g) = 20 ng
g to bp const = 1.096 x 10^-21 g
genome size = 4.6 x 10^6 bp

20 x 10^-9 g / (1.096 x 10^-21 g/bp x 4.6 x 10^6 bp) = 4.0 x 10^6 copies