User:Tara K. Luckau/Notebook/Team ConGen/2010/10/26

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Contents

GelRed Info

Gel Test

Cast Gel

  • small caster: 10cm x 10cm x 0.5cm = 50mL TAE
  • 50mL TAE + 1g agarose in 250mL Erlenmyer flask
  • small stirbar, hotplate set at 200, stir set at 150 for about 10 min, until clear
  • let cool (about 5 min)
  • add 5µL GelRed 10,000X, swirl
  • pour into caster, set comb
  • let set (about 5min)

Load Gel

  • 4µL Tara Ladder (T)
  • 4µL Justin Ladder (J)
  • 2µL Sample (H6) (added minute after applying current)

Run Gel

  • 110V, 20 min
  • 150V, 10 min

Image Gel

  • Ladders show!
    both Tara's and Justin's
  • Sample is very faint, may be due to smaller volume placed
  • ~200-300bp fragment
  • (Lance et al 2009, says Scun2 has amplicon length of 131-191)
  • do another gel with subset of PCR product, full volume



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