Femto™ Human DNA Quantification Kit

Quantify as little 20 femtograms of human DNA in as little as 1 µl of sample.
High specificity and sensitivity for human DNA in a background of non-target DNA.
Fast and simple: add samples to the PreMix... and quantify.

Product Size Catalog # Price Qty
Femto™ Human DNA Quantification Kit 100 Rxns E2005
$209.00

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.

Reagent Preparation:

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) 

 

  1. Aliquot 18 µl of the Femto Human qPCR Premix into each well planned for use. 3

 

  1. Add 2 µl of Human DNA Standards (#1-7) into the appropriate wells.  Remember to change pipette tips after the addition of each Human DNA Standard to a well.

 

  1. Add 1 to 3 µl of each Unknown Test Sample to the appropriate wells containing the Master Mix.  Remember to change pipette tips after the addition of each Unknown Test Sample.  DO NOT ADD Unknown Test Samples to wells containing Human DNA Standards or No Template Control.

 

  1. Add 2 µl of No Template Control (#8) into the appropriate wells.  Remember to change pipette tips after addition of each No Template Control volume.

 

  1. Seal the qPCR plate with an optically transparent sealing film or qPCR tube strips with tube strip caps that are compatible with the real-time/quantitative PCR instrument being used.

 

  1. Centrifuge the qPCR plate or qPCR tube strips to eliminate bubbles and to bring any droplets to the bottom of the well.

 

 

Thermocycling Parameters:

                                                            Temperature               Time 

-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

 

Analysis:

 

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.    

 

Human DNA Standards

Amount of Human DNA Input (ng)/ Reaction Well

Standard 1

20

Standard 2

2

Standard 3

0.2

Standard 4

0.02

Standard 5

0.002

Standard 6

0.0002

Standard 7

0.00002

 

 

 

 

 

 

 

 

 

 

 

 

For specific notes and additional information, please see the product protocol PDF.
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