1X dsDNA HS Assay Kit

Description

Laboratories measuring low concentrations of double-stranded DNA may find that UV absorbance at 260 nm is affected by other substances in the sample. Preparing a working solution before every assay also adds a handling step.

The Biofargo 1× dsDNA HS Assay Kit provides a ready-to-use, fluorescence-based method for high-sensitivity dsDNA quantification. The supplied 1× dsDNA HS Working Solution requires no dilution. Add the sample, incubate for 2 minutes at room temperature protected from light, and measure the fluorescence.

The assay is highly selective for dsDNA over RNA. It accepts 1–20 µL of sample in a final assay volume of 200 µL. The current product details state an initial dsDNA concentration range of 10 pg/µL–100 ng/µL, depending on sample volume, and a total assay range of 0.2–100 ng.

The kit supports Qubit 2, Qubit 3, Qubit 4, and Qubit Flex fluorometers. It can also be used with a fluorescence microplate reader at 485 nm excitation and 530 nm emission. When protected from light, the fluorescent signal remains stable for up to 3 hours after incubation.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Specifications

Features

• Ready-to-use formulation: The supplied 1× dsDNA HS Working Solution can be used without dilution.

• High-sensitivity dsDNA quantification: The current product details state a total assay range of 0.2–100 ng.

• Selective dsDNA measurement: The assay is highly selective for double-stranded DNA over RNA.

• Flexible sample input: Each assay accepts 1–20 µL of sample in a final volume of 200 µL.

• Short incubation: Incubate for 2 minutes at room temperature while protected from light.

• Stable reading window: The fluorescent signal remains stable for up to 3 hours when protected from light.

• Multiple instrument formats: The kit supports the specified Qubit fluorometers and a fluorescence microplate reader.

• Contaminant tolerance: DNA standards were evaluated with BSA, sodium acetate, ammonium acetate, sodium chloride, magnesium chloride, phenol, chloroform, ethanol, SDS, Triton X-100, ssDNA, polyethylene glycol, RNA, and dNTPs. In the supplied results, “OK” usually represents less than 10% perturbation at the indicated final assay concentration.

Applications

• High-Sensitivity dsDNA Quantification with a Qubit Fluorometer: Equilibrate the kit components to room temperature. Use thin-wall, clear 0.5 mL PCR tubes and label their lids rather than their sides. Prepare each standard with 190 µL of 1× dsDNA HS Working Solution and 10 µL of QS0 or QS1. Prepare each sample with 180–199 µL of Working Solution and 1–20 µL of sample, for a final volume of 200 µL. Use a 2 µL pipette when adding 1–2 µL of sample. Vortex each tube for 3–5 seconds, then incubate for 2 minutes at room temperature protected from light. On a Qubit 4 Fluorometer, select dsDNA, then 1× dsDNA High Sensitivity. Read QS0 followed by QS1, select the sample volume and output units, and read the samples. Avoid warming the tubes by holding them before measurement. Do not use calibration curves generated with other kits.

• High-Sensitivity dsDNA Quantification with a Fluorescence Microplate Reader: Equilibrate the kit components to room temperature and mix thoroughly. Use a black-wall, clear-bottom 96-well microplate. Add 190 µL of Working Solution to each standard well and 180–199 µL to each sample well. Serially dilute QS1 to prepare standards from 0 to 10 ng/µL, then add 10 µL of each standard to its designated well. Add 1–20 µL of each sample so every well contains exactly 200 µL. Vortex the plate for 10–15 seconds and incubate for 2 minutes at room temperature protected from light. Read at 485 nm excitation and 530 nm emission. Generate a standard curve to calculate unknown concentrations. If a sample exceeds the linear range, dilute it and repeat the assay.

Kit Components

Storage & Shipping

• Store at 2–8°C, protected from light.

• Ship on ice.

Frequently Asked Questions (FAQs)

Documents

Product User Manual