Difference between revisions of "Pouring an Agarose Gel"

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# Weigh out agarose in 125 ml flask (eg. 0.35 grams agarose for 1% gel).
 
# Weigh out agarose in 125 ml flask (eg. 0.35 grams agarose for 1% gel).
# Add 35 mL of TAE buffer
+
# Add 35 mL of 1X TAE buffer
# Microwave for 1 minute
+
# Microwave for 50 seconds
# Swirl and microwave for 10 seconds
+
# Swirl and microwave for 5 seconds - repeat 3 times
# Let flask cool enough to touch, add 5 µL of 10 mg/ml Ethidium Bromide (WEAR GLOVES)
+
# Place flask in 55 C water bath to cool
 +
# Add 5 µL of 10 mg/ml Ethidium Bromide (CAUTION: WEAR GLOVES)
 
# Swirl to mix and pour into gel tray with comb
 
# Swirl to mix and pour into gel tray with comb

Revision as of 17:01, 15 February 2017

Pouring an Agarose Gel

  1. Go to http://www.bio.davidson.edu/courses/Molbio/Protocols/molwt.html to determine the percentage of agarose and calculate amount of agarose for 35 ml.
  1. Weigh out agarose in 125 ml flask (eg. 0.35 grams agarose for 1% gel).
  2. Add 35 mL of 1X TAE buffer
  3. Microwave for 50 seconds
  4. Swirl and microwave for 5 seconds - repeat 3 times
  5. Place flask in 55 C water bath to cool
  6. Add 5 µL of 10 mg/ml Ethidium Bromide (CAUTION: WEAR GLOVES)
  7. Swirl to mix and pour into gel tray with comb