About 95% of the GlycoBlue Coprecipitant will be removed from the recovered RNA by precipitation with 2.5 M Lithium Chloride (LiCl). (Note: no alcohol is used in this precipitation – simply bring the sample to 2.5 M LiCl, and spin to pellet.

How do you remove GlycoBlue?

About 95% of the GlycoBlue Coprecipitant will be removed from the recovered RNA by precipitation with 2.5 M Lithium Chloride (LiCl). (Note: no alcohol is used in this precipitation – simply bring the sample to 2.5 M LiCl, and spin to pellet.

How much GlycoBlue do I add?

By using 1-2 ul per one RNA isolation, one GlycoBlue vial should be sufficient for 150-300 isolations adding an extra 15-30 cents to each reaction.

How does glycol blue help with alcohol precipitation of DNA?

When a typical acetate/alcohol precipitation is done, the GlycoBlue Coprecipitant will precipitate with the nucleic acids, facilitating good RNA or DNA recovery while increasing the size and visibility of the pellet.

How do you precipitate DNA with sodium acetate?

1. Add: 0.1 vols 3M Sodium acetate 2.5-3 vols ice cold 100% Ethanol Vortex to mix thoroughly. 2. Precipitate at -200C for 1 hour or overnight or -80 0C 1 hr (overnight will give more precipitation if RNA amount is low) 3.

How do you precipitate RNA?

After the salt concentration has been adjusted, the RNA may be precipitated by adding 2.5 volumes of ethanol or 1 volume of isopropanol and mixing thoroughly, followed by chilling for at least 15 minutes at -20° C.

How do you precipitate DNA with ethanol?

Ethanol precipitation of DNA:

  1. Add 2 volumes of ethanol to the sample and freeze at –20°C for at least 1 hour or overnight for best results.
  2. Centrifuge the sample at full speed for 20 minutes to collect all material.
  3. Wash with 70% ethanol, then centrifuge for 10–15 minutes to pellet the DNA.

Why is alcohol used to precipitate the DNA?

The main role of monovalent cations and ethanol is to eliminate the solvation shell that surrounds the DNA, thus allowing the DNA to precipitate in pellet form. Additionally, ethanol helps to promote DNA aggregation.

Why is isopropanol used in DNA extraction?

7. Isopropanol/ethanol. Alcohol is used to precipitate the DNA out of the extraction solution, so we can wash all those salts and chemicals away and then dissolve it in our final solvent—usually water or some variant of Tris-EDTA solution.

Why is sodium acetate used in DNA precipitation?

The Role of Salt… A commonly used salt is sodium acetate. In solution, sodium acetate breaks up into Na+ and [CH3COO]–. The positively charged sodium ions neutralize the negative charge on the PO3– groups on the nucleic acids, making the molecule far less hydrophilic and, therefore, much less soluble in water.

Does sodium acetate precipitate?

No, it does not.

How do I perform a DNA precipitation to concentrate my sample?

FAQ

  1. Add 1/10 volume of 3 M Na-Acetate pH 5.2, and 2 to 2.5 volumes of ice-cold 100% ethanol to the DNA sample.
  2. Mix, and store at –20°C for at least 1 h to precipitate the DNA.
  3. Recover the precipitated DNA by centrifugation at full speed in a microcentrifuge for 15–20 min.

What is the best solution for storage of RNA?

RNA may be stored in a number of ways. For short-term storage, RNase-free H2O ( with 0.1 mM EDTA) or TE buffer (10 mM Tris, 1mM EDTA) may be used. RNA is generally stable at -80° C for up to a year without degradation.

Can glycoblue™ coprecipitant be added to nucleic acid solutions?

GlycoBlue™ Coprecipitant can be added to nucleic acid solutions at a final concentration of 50–150 µg/mL. When a typical acetate/alcohol precipitation is done, the GlycoBlue™ Coprecipitant will precipitate with the nucleic acids, facilitating good RNA or DNA recovery while increasing the size and visibility of the pellet.

Can glycoblue be used to precipitate RNA?

Indeed, if you used a high amount of GlycoBlue to precipitate your RNA, it might be difficult to get rid of the dye despite all the washes, and this could affect all QC and concentrations measurements based on spectroscopy (and maybe even fluoresence).

How does the glycoblue coprecipitant work?

When a typical acetate/alcohol precipitation is done, the GlycoBlue Coprecipitant will precipitate with the nucleic acids, facilitating good RNA or DNA recovery while increasing the size and visibility of the pellet.

Does glycoblue affect NanoDrop results?

Just a warning, Glycoblue alone has peaks you will see on the Nanodrop at BOTH 230 and 260 that will effect results if your nucleic acid concentration is low enough and you’ve concnetrated the Glycoblue enough. For instance, 2 ml down to 20 yielding 100ng/ul, you will see the dye a little.