Agarose Gel Calculator

Turns a gel volume and target percentage into agarose, buffer and stain volumes, and points at the percentage that resolves the fragment size you need.Enter the gel volume and target percentage and the recipe appears. Casting several gels? Raise the count.

Stain fold and fragment size are optional. Leave them empty and only those lines drop out.

Gel percentage and separation range

Agarose concentration (% w/v)Separation range (kb)
0.3*560
0.5130
0.70.812
1.00.510
1.20.47
1.50.23
2.0*0.12

The source marks the 0.3% and 2.0% rows with an asterisk (*), but prints no footnote for it anywhere in the document. The mark is carried across as-is rather than explained.

Source: QIAGEN Bench Guide, “DNA analysis using analytical gels”, table “Concentration of agarose used for separating fragments of different sizes”.

How to use

  1. Enter the volume of one gel in mL and the target percentage. Percentage here is weight per volume, so 1% means 1 g of agarose in 100 mL of buffer.
  2. Casting more than one gel? Raise the count — volume and mass scale together.
  3. Pick the running buffer stock you keep. 50× TAE, 10× TBE and 5× TBE are set up; for anything else enter the fold yourself.
  4. Enter the fold of your stain stock and its volume appears too. Stain comes out of the water rather than on top of it, so the total stays the volume you asked for.
  5. Enter the fragment size in bp and the table below marks every percentage that resolves it.

Formula and practical notes

An agarose gel sieves DNA by pore size. More agarose means tighter pores and better separation of small fragments; less agarose opens the pores for large ones. That is why “what percentage do I need” is answered by a table, not by a formula.

The arithmetic

Four lines, and total volume is the volume of one gel times the number of gels.

Agarose (g)
target % ÷ 100 × total volume (mL)
Buffer stock (mL)
total volume ÷ stock fold
Stain (μL)
total volume ÷ stain fold × 1000
Water (mL)
total volume − buffer stock − stain

Practical notes

  • Ethidium bromide (EtBr) is a suspected mutagen. Wear gloves, and treat the gel, the running buffer, the comb and the tray as contaminated. Casting it into the gel is convenient but contaminates the tank and slows linear DNA by about 15%; staining after the run uses less and keeps the tank clean.
  • The concentration usually quoted for EtBr is a final 0.5 μg/mL. This calculator deliberately ships no stain as a default — enter the fold printed on the bottle you have.
  • TAE has a lower buffering capacity than TBE and is exhausted sooner on a long run. TBE gives sharper bands and is particularly recommended below 1 kb, but its borate ions complex with sugars and make DNA harder to recover from the gel. For a gel you intend to cut and purify, TAE is the easier choice.
  • Use ultrapure agarose. Polysaccharides, salts and proteins all distort migration, and the effect grows with the percentage.
  • This is arithmetic on volumes and masses. What a gel actually resolves also depends on voltage, run time and the rig, so treat the table ranges as a starting point.

Frequently asked questions