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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