About Femto™ Human DNA Quantification Kit
The Femto™ Human DNA Quantification Kit can detect and quantify human DNA with high specificity and sensitivity. Human DNA can be reliably quantified in a background of non-human DNA such as bacterial, fungal, animal, plant DNA, etc. This is essential for downstream applications that require accurate DNA input amounts such as STR analysis. With the Femto™ Human DNA Quantification Kit, dependably quantify as little as 20 fg from 1 µl of purified biological liquids, anthropological samples, or forensic DNA samples.
|Sample Sources||Detect and quantify high quality human 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 less than 1 copy of human gDNA.|
Reliable standards for the quantification of human DNA: Human DNA Standards performed in duplicates 10-fold serial dilutions ranging from 20 ng to 20 fg.
Specificity: Amplification plots demonstrate the specificity of the Femto™ Human DNA Quantification Kit. Bovine gDNA, porcine gDNA, and murine gDNA exhibited delayed amplification similar to the No Template Control (black) (≥35 cycles). 0.2 ng of human brain gDNA (orange), human gDNA extracted from urine (gold), bovine gDNA (red), porcine gDNA (light blue), and murine gDNA (purple) were used.
✓It is recommended that all reagents and qPCR reactions be prepared using clean techniques to prevent contamination. 1
✓Femto™ Human 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™Human qPCR Premix.
✓Femto™ Human qPCR Premix should be protected from direct light exposure. Minimize freeze-thaw cycles.
✓ Human 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 Human DNA Quantification:
Aliquoting Femto™ Human 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 59 ˚C 30 seconds 40 cycles 4
-Extension 72 ˚C 1 minute
-Final Extension 5 72 ˚C 7 minutes
Use the Human 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 human DNA, Standard 2 reaction wells contain 2 ng of human DNA, etc.