Team:Pasteur Paris/Test

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Week 29 16 - 22 July
Week 30 23 - 29 July
Week 31 30 July - 5 Aug
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Agarose Gel Preparation


Aim

Prepare an 8% agarose gel for the electrophoresis of DNA samples.


Materials

  • UltraPure™ Agarose (Invitrogen, 16500-100)
  • UltraPure™ 10X TAE Buffer (Invitrogen, 15558)
  • Gel Green Nucleic Acid Stain (Biotium, 41005)
  • Scale
  • Microwave
  • Spatula
  • Erlenmeyer (250 mL)
  • Measuring cylinder
  • PowerPac™ Basic (Bio-Rad, 1645050)
  • Owl Scientific (Thermofisher) B2 Series gel block (plus Biorad powerpack adaptor)

Procedure


  1. Prepare 600 mL of TAE 1X by diluting 60 mL of 10X buffer in 540 mL of deionized water.
  2. Weigh 0.6 g of agarose on a scale.
  3. Place the agarose in an Erlenmeyer.
  4. Fill the Erlenmeyer with 75 mL of TAE 1X.
  5. Heat the Erlenmeyer for 2 min 30 s at 350W in the microwave oven.
  6. Stir and place it again in the microwave for an additional minute.
  7. Let the mixture cool down to around when it just warm to touc and add 5 μL of Gel Green.
  8. Pour the agarose in the horizontal electrophoresis system. Don’t forget to place the comb before!
  9. Let the gel cool down for 20-30 minutes before deposing the samples.


Salut