Variants:

 

  1. 1. Depend on type of molecules:

 

For ds DNA can be used kits below:

Quant-iT™ PicoGreen ® dsDNA Assay  Kit (Invitrogen P7589);

Qubit® dsDNA Broad Range Assay Kit (Invitrogen Q32850);

Qubit® dsDNA High Sensitivity Assay  Kit (Invitrogen Q32854).

For ss DNA  -  Qubit® ssDNA Assay Kit (Invitrogen Q10212).

For RNA  Qubit® RNA Assay Kit (Invitrogen Q10212).

 

  1. 2. Depend on type of cell type to measure fluorescence:

 

The following products can be used with the Qubit® 2.0 Fluorometer: Qubit® Assay Tubes 500 tubes Q32856  or thin wall, clear 0.5 ml tubes (Axygen PCR-05-C), the volume of measured sample -  200 mkl.

QuantiFluor (Promega)  - reusable methacrylate cuvette section of 10x10 mm, the volume of the measured sample 2 ml

 

Features sensitivity of diffrent variations of the method:

 

Assay

The range of measured concentrations

Quant-iT™ PicoGreen ® dsDNA Assay  Kit

2.5 ng/mkl – 100 mg/ml

Qubit® dsDNA BR Assay Kit

10 ng/mkl – 10000 ng/ml

Qubit® dsDNA HS Assay  Kit

0.5 ng/mkl – 500 ng/ml

Qubit® ssDNA Assay  Kit

1 ng/mkl – 1000 ng/ml

Qubit® RNA Assay  Kit

20 ng/mkl – 1000 ng/ml

 

Protocol:

  1. 1. To prepare working solution of fluorescent due.
  2. 2. To calibrate fluorometer with appropriate standard solutions according with the user guide..
  3. 3. To carry out measurements. If the concentration of the resulting value is outside the measurement set or device to perform the necessary dilution of the sample and repeat the measurement.

References:

  1. 1. Quant-iT™ PicoGreen ® dsDNA Reagent and Kits. Invitrogen. 2008

(http://tools.thermofisher.com/content/sfs/manuals/mp07581.pdf)

  1. 2. QuantiFluor™ Handheld Fluorometers Operating Instructions. Technical manual. Promega Corporation. 2011.

(https://www.promega.com/~/media/files/resources/protocols/technical%20manuals/101/quantifluor%20handheld%20fluorometers%20protocol.pdf)

  1. 3. Qubit® dsDNA BR Assay Kits. Manual. Thermo Fisher Scientific Inc. 2015.

(https://tools.thermofisher.com/content/sfs/manuals/Qubit_dsDNA_BR_Assay_UG.pdf)

  1. 4. Qubit™ dsDNA HS Assay Kits. Manual. Thermo Fisher Scientific Inc. 2015.

(https://tools.thermofisher.com/content/sfs/manuals/Qubit_dsDNA_HS_Assay_UG.pdf)

  1. 5. Qubit™ ssDNA Assay Kits. Manual. Thermo Fisher Scientific Inc. 2015.

(https://tools.thermofisher.com/content/sfs/manuals/Qubit_ssDNA_Assay_UG.pdf)

  1. 6. Qubit™ RNA Assay Kits. Manual. Thermo Fisher Scientific Inc. 2015.

(https://tools.thermofisher.com/content/sfs/manuals/Qubit_RNA_BR_Assay_UG.pdf)