What is PicoGreen?

PicoGreen is a fluorescent probe that binds dsDNA and forms a highly luminescent complex when compared to the free dye in solution. On binding to DNA, intercalation and electrostatic interactions immobilize the dye molecule, resulting in a >1000-fold enhancement in its fluorescence.

What is a PicoGreen DNA assay?

PicoGreen dsDNA quantitation reagent is a highly sensitive fluorescent nucleic acid stain which is used to quantitate double-stranded DNA. PicoGreen is widely used in molecular biological procedures such as DNA fragment purification for subcloning, cDNA synthesis for library production as well as primer assays.

Does qubit use PicoGreen?

Samples processed using the Quant-iT PicoGreen dsDNA Assay Kit are read with a standard fluorescence microplate reader, and the data are plotted with a graphics software package of the user’s choice. Qubit dsDNA assay kits are convenient for fewer samples, since the samples are read one at a time.

What is OliGreen?

Quant-iT™ OliGreen® oligonucleotide reagent is an ultra-sensitive fluorescent nucleic acid stain. Short, synthetic oligonucleotides are used in a number of molecular biology techniques, including: DNA sequencing, site- directed mutagenesis, DNA amplification, and in situ hybridization.

How do you make a PicoGreen assay?

1) Prepare 40 mL of 1X TE buffer by adding 2 mL of 20X TE buffer to 38 mL of ultrapure distilled water. 2) Dilute enough PicoGreen reagent for the assay 200 times in 1X TE buffer….

1. Prepare and aliquot reagents.
2. Digest samples with Papain and equalize their volumes for the assay.
3. Perform the PicoGreen Assay.

How does DNA fluorescence work?

Fluorescent labels. Fluorescent dyes have been used for decades to stain biomolecules for imaging and detection applications. Many of these dyes bind to DNA and RNA, causing the nucleic acids to become fluorescent and therefore readily detected in a fluorescence microscope, a flow cytometer or a gel.

How does DNA assay work?

Uses and Advantages. The assay can be used to detect and quantify many types of RNA or DNA target. In the assay, branched DNA is mixed with a sample to be tested. Thus, small amounts of a nucleic acid can be detected and quantified without a reverse transcription step (in the case of RNA) and/or PCR.

How can nucleic acids be Quantitated?

One of the more commonly used practices to quantitate DNA or RNA is the use of spectrophotometric analysis using a spectrophotometer. A spectrophotometer is able to determine the average concentrations of the nucleic acids DNA or RNA present in a mixture, as well as their purity.

What is the most accurate way to quantify DNA?

Choosing the Best DNA Quantification Method

1. Real-Time Polymerase Chain Reaction. Real-time PCR, sometimes referred to as qPCR, is one of the most common RNA and DNA quantification methods in use today, owing to its sensitivity, specificity, and dynamic range.
2. UV-Vis Spectrophotometry.
3. Fluorescence.

What reagent is used for quantifying DNA?

Fluorescence detection with Hoechst 33258 reagent is a sensitive and precise method for DNA quantification if the G+C content is less than 50%. In addition, this method allows quantification of very low levels of DNA (nanogram scale).

How does a DNA assay work?

The assay can be used to detect and quantify many types of RNA or DNA target. In the assay, branched DNA is mixed with a sample to be tested. The detection is done using a non-radioactive method and does not require preamplification of the nucleic acid to be detected. The assay entirely relies on hybridization.

What is FISH lab?

Fluorescence in situ hybridization (FISH) is a laboratory technique for detecting and locating a specific DNA sequence on a chromosome. The technique relies on exposing chromosomes to a small DNA sequence called a probe that has a fluorescent molecule attached to it.