We believe this site might serve you best:

United States

United States

Language: English

Promega's Cookie Policy

Our website uses functional cookies that do not collect any personal information or track your browsing activity. When you select your country, you agree that we can place these functional cookies on your device.

What percentage agarose is needed to sufficiently resolve my DNA sample?

SEARCH ARTICLES

Related Resources

LabFact #16

More restriction enzyme is generally required to digest supercoiled DNA than linear DNA. If you linearize supercoiled DNA with one restriction enzyme, less of a second restriction enzyme is required for complete digestion.

  • Print
  • Email

Linear DNA can be resolved by size using agarose gels of various concentrations. The greater the percentage of agarose, the smaller the linear DNA that can be resolved. The sugar polymers that make up the agarose gel matrix (powdered agarose heated in appropriate buffer, poured into a gel tray and allowed to solidify) act like a sieve. The greater the agarose concentration, the smaller the pores created in the gel matrix, and the more difficult it is for large linear DNA molecules to move through the matrix. Changing the agarose concentration changes the size of the sieve matrix of the gel. However, there is an upper and lower limit to accurate separation of DNA molecules using agarose gel electrophoresis. To estimate the size of a linear DNA fragment and to diagnose any electrophoresis issues, run at least one DNA molecular weight marker [e.g., BenchTop 1kb DNA Ladder (Cat.# G7541)] along with the experimental sample. A table highlighting the various agarose percentages and molecule sizes that can be separated is shown below.

Resolution of Linear DNA on Agarose Gels.

Recommended % Agarose Optimum Resolution for Linear DNA
0.5 1,000–30,000bp
0.7 800–12,000bp
1.0 500–10,000bp
1.2 400–7,000bp
1.5 200–3,000bp
2.0 50–2,000bp

For standard analytical gels, we recommend an agarose like Agarose, LE, Analytical Grade (Cat.# V3121). For preparative gels, where fragments are to be separated and cut from a gel, a low-melting-point agarose such as Agarose, LMP, Preparative Grade for Large Fragments (>1,000bp; Cat.# V2831) or Agarose, LMP, Preparative Grade for Small Fragments (10 to 1,000bp; Cat.# V3841) is advantageous.

Products may be covered by pending or issued patents or may have certain limitations. Please visit our Web site for more information.

Choose your country

Americas

Brazil
Canada
United States

Pacific Asia

Australia
India
Japan
Korea, Republic of
Singapore

Europe

Austria
Belgium
Denmark
Estonia
Finland
France
Germany
Iceland
Italy
Luxembourg
Netherlands
Norway
Poland
Spain
Sweden
Switzerland
United Kingdom