About Femto™ Bacterial DNA Quantification Kit
The Femto™ Bacterial DNA Quantification Kit can 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, plant DNA, etc. This is essential for downstream applications that require accurate DNA input amounts such as quantifying bacteria DNA template for Next-generation sequencing library preparation and metagenomic analysis. With the Femto™ Bacterial DNA Quantification Kit, dependably quantify as little as 20 fg of bacterial DNA from 1 µl of purified biological liquids, bacterial cultures, or environmental DNA samples.
|Sample Sources||Detect and quantify high quality bacterial DNA from any purified mixed DNA sample.|
|Compatibility||Product is designed to be compatible with any real-time and quantitative PCR instrument.|
|Bacterial DNA Detection and Quantification||Detection range of 20 fg-20 ng from as little as 1 µl of sample. The kit can be used to detect down to 5 copies of Escherichia coli genomic DNA.|
Reliable standards for the quantification of bacterial DNA: Bacterial DNA Standards (measured in duplicates) comprise a 10-fold dilution series ranging from 20 ng to 20 fg.
Amplification of bacterial DNA from a variety of samples: Amplification plots of the Femto™ Bacterial DNA Quantification Kit of inputs of purified DNA extracted from different sources are shown: Chinchilla feces (purple), sludge (light blue), soil (orange), and E. coli culture (pink). Bacterial DNA Standards (green) and No Template Control (black) are also shown.
✓It is recommended that all reagents and qPCR reactions be prepared using clean techniques to prevent contamination. 1
✓Femto™ Bacterial qPCR Premix should be completely thawed at room temperature, mixed by flicking the tube, centrifuged briefly, and then placed on ice. DO NOT VORTEX Femto™ Bacterial qPCR Premix.
✓Femto™ Bacterial qPCR Premix should be protected from direct light exposure. Minimize freeze-thaw cycles.
✓Bacterial DNA Standards (#1-7) should be completely thawed at room temperature, mixed by vortexing, centrifuged briefly, and then placed on ice. 2
✓All reagents should be kept on ice immediately after thawing.
Protocol for Bacterial DNA Quantification:
Aliquoting Femto™ Bacterial qPCR Premix and qPCR Set Up (for qPCR tube strips or qPCR plates)
-Initial Denaturation 95 ˚C 10 minutes
-Denaturation 95 ˚C 30 seconds
-Annealing 50 ˚C 30 seconds 40 cycles 4
-Extension 72 ˚C 1 minute
-Final Extension 5 72 ˚C 7 minutes
Use the Bacterial DNA Standards table below to generate a standard curve to quantify Unknown Test Samples. For example, the Standard 1 reaction wells contain 20 ng of bacterial DNA, Standard 2 reaction wells contain 2 ng of bacterial DNA, etc.