- Quantify as little 20 femtograms of bacterial DNA in as little as 1 µl of sample.
- High specificity and sensitivity for bacterial DNA in a background of non-target DNA.
- Fast and simple: add samples to the PreMix… and quantify.
The Femto Bacterial DNA Quantification Kit can be used to detect and quantify bacterial DNA with high specificity and sensitivity. Bacterial DNA can be reliably quantified in a background of non-bacterial DNA such as fungal, animal, and plant DNA, etc. This is essential for downstream applications that require accurate DNA input including quantifying bacteria DNA template for Next-generation sequencing library preparation and metagenomic analysis amongst others. With the Femto Bacterial DNA Quantification Kit, one can dependably quantify as little as 20 fg DNA from 1 µl purified from biological liquids, bacterial cultures, or environmental DNA samples.
|Detection Range||20 fg to 20 ng DNA|
|Equipment||Real-time quantitative PCR system,vortex, microcentrifuge, PCR tube strip or PCR plate, optically transparent sealing film for PCR plate or tube strip caps|
|Sample Source||Purified DNA|
Q1: Is there any general advice on GLP for qPCR?
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.
Q2: Does qPCR Premix contain ROX?
Our qPCR Premixes does not contain ROX Reference Dye. Ensure that the ROX channel of your device is switched off.
Q3: Which settings can be used for signal detection?
Our qPCR Premixes contains Syto9. You can use the SYBR Green channel for signal detection.
Q4: Are the Femto kits certified for low bioburden?
No, kits are not certified.
Q5: What is the origin of the bacterial DNA Standards?
-Bacterial Standards are generated from E. coli strain JM109.
-Standards are QC tested serial dilutions of genomic DNA.
Q6: What is strain JM109?
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). Ref: https://www.ncbi.nlm.nih.gov/pubmed/9278503
Q7: How is the copy number of the 16S rRNA gene calculated?
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.
Q8: How to calculate the amount of genomic copies detected with E2006?
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
Femto Bacterial DNA Quantification Kit