What is PicoGreen used for?
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. The most widely used method of measuring nucleic acid concentration is the determination of absorbance at 260nm (A260).
How does PicoGreen bind to DNA?
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 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.
What does PicoGreen measure?
PicoGreen dsDNA Quantitation Reagent is an. ultra-sensitive fluorescent nucleic acid stain for. quantitating double-stranded DNA (dsDNA) in. molecular biological procedures such as cDNA. synthesis for library production and DNA.
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….
- Prepare and aliquot reagents.
- Digest samples with Papain and equalize their volumes for the assay.
- Perform the PicoGreen Assay.
What does PicoGreen stain?
The PicoGreen® dye is a fluorescent nucleic acid stain for quantitating double-stranded DNA (dsDNA). The main disadvantage of general UV spectroscopy for dsDNA quantitation is the contribution of signal from single- stranded DNA (ssDNA) and other contaminants, such as protein and extraction buffers.
Is qubit or Nanodrop more accurate?
While spectroscopic methods like UV/VIS (eg. Nanodrop spectrophotometer) provide adequate estimation at concentrations higher than 10 ng/µL, fluorometric quantification (eg. qubit fluorometer) will always be more accurate and specific (measuring DNA only).
What is a 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 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 kind of Kit do you need for Picogreen?
Quant-iT™ PicoGreen ® dsDNA Kit (Invitrogen, #P11496). This kit contains Quant-iT™ PicoGreen ® dsDNA Reagent (10 vials, each containing 100 µL aliquots) Black, flat-bottomed 96-well plates (e.g. Dynex, #3010 or Greiner Cellstar, #655086)
What can the Picogreen assay be used for?
The PicoGreen assay is useful for quantifying the amount of double-stranded DNA (dsDNA) that is present in a given sample. The measured amount of dsDNA can then be used as a marker for cell proliferation.
What is the use of Picogreen dsDNA reagent?
Applications: The PicoGreen® dsDNA Quantitation Reagent and Kits are ideal for PCR-based assays, microarray samples, DNA damage assays, enzyme activity assays, genomic DNA quantitation, measuring dsDNA in complex mixtures, and viral DNA quantitation. For Research Use Only. Not for use in diagnostic procedures.