DNA, RNA and protein quantitation technique

Variants:

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).

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. To prepare working solution of fluorescent due.
2. To calibrate fluorometer with appropriate standard solutions according with the user guide..
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: