Quantifying DNA in Solution with PicoGreen
Hello everyone,
Today I am discussing a different method for quantification of DNA ie Quantifying DNA in Solution with PicoGreen
I have covered similar topics earlier....
DNA QUANTIFICATION USING MINIGELS
Estimating the concentration of DNA by Fluorometry using Hoechst 33258
PicoGreen dsDNAQuantitation 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 fragment purification for subcloning, as well as diagnostic applications, such as quantitating DNA amplification products and primer extension assays.
The PicoGreen assay allowed the detection of 25 pg/ml dsDNA , surpassing the sensitivity achieved with Hoechst 33258 by 400-fold. The assay is most quickly and easily performed using a handheld fluorometer.
The standard PicoGreen assay protocol is also simpler than the Hoechst 33258 method, because a single concentration of the PicoGreen Reagent allows detection over the full dynamic range of the assay.Unlike hoechst – based assay, assay using a single concentration of picogreen are linear over four orders of magnitude in DNA concentration from 1ng/ml to 1 µg/mL.
PRINCIPLE:-
PicoGreen dsDNA Quantitation Reagent is an ultra-sensitive fluorescent nucleic acid stain for quantitating double-stranded DNA (dsDNA) in molecular biology procedures.The brightness of this reagent is due to its high quantum yield and large molar extinction coefficient. These procedures include cDNA synthesis for library production, DNA fragment purification for sub cloning, and diagnostic applications, such as quantitating DNA amplification products, and primer extension assays.
REAGENTS
Picogreen dsDNA quantitation reagent
Reference DNA (eg calf thymus DNA) at concentration of 2 µg/mL in TE(pH 7.5)
TE (pH 7.5)
To read more http://microamaze.blogspot.in/2016/01/quantifying-dna-in-solution-with.html
Other methods for DNA Quantification DNA QUANTIFICATION USING MINIGELS Estimating the concentration of DNA by Fluorometry using Hoechst 33258 Extraction and Purification of Plasmid DNA by Alkaline Lysis MethodEstimation of DNA by Diphenylamine method
With love
- Dixy