How to make agarose gels for DNA separation

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How to make agarose gels for DNA separation 

Protocol for 1% agarose gel

 

  • Weigh in 1g of agarose in an erlenmeyer flask or glass bottle
  • Add 99 ml of 1x TAE buffer 
  • Heat in microwave until the agarose dissolves completely (shake) 
  • Cool down agarose with tap water until flask can be touched with bear hands 
  • Add DNA dye (for example midori green by Nippon Genetics) 1:1000 e.g. 1µl; shake 
  • Pour gel into gel chamber 
  • Let solidify for 30 min at RT 
  • Take gel, place into agarose gel electrophoresis chamber, remove comb gently 
  • Add 1x TAE buffer and cover gel with buffer completely, make sure that slots are filled with buffer 
  • Prepare DNA samples: For example if you have 20 µl of sample, add 4 µl of 6x loading dye 
  • Load DNA marker (GeneRuler 1kb 0.5µg/µl  by Thermo Fisher) and samples into gel slots 
  • Run the gel at 90 Volts for 45 min to 1 hour. 
  • Analyse DNA under UV light or imager 
50x TAE Buffer :

Component

Concentration

Tris base

2 M

Disodium EDTA

50 mM

Acetic Acid

1 M


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