Femto™ Bacterial DNA Quantification Kit

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.

Product Size Catalog # Price Qty
Femto™ Bacterial DNA Quantification Kit 100 Rxns E2006
$209.00

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.

Reagent Preparation:

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) 

 

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

 

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

 

Thermocycling Parameters:

                                                            Temperature               Time 

-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

 

Analysis:

 

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.    

 

Bacterial DNA Standards

Amount of Bacterial 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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