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

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GelRed Info

 * Yesterday, used 27µL 3x Gel Red into warm agarose/TAE mix (precast)
 * Biotium's Product Information sheet on GelRed: [[Media:Biotium_GelRedProductProtocol.pdf‎]]

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)
 * [[Image:20101026 GelSchematic.jpg|400 px]]

Run Gel

 * 110V, 20 min
 * 150V, 10 min

Image Gel

 * Ladders show!
 * both Tara's and Justin's
 * [[Image:20101026_Full.TIF|300 px]]
 * 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)
 * [[Image:20101026_Zoom.TIF|300 px]]
 * do another gel with subset of PCR product, full volume


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