User:Stuart McKellar/Notebook/Bird Sex Testing/2012/09/11

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Notes from last time

  • I used a 5x buffer for the agar gel whoops
  • I needed to set the power supply for the electrophoresis tank to 200V and 80mA minimum*
  • DNA needs to run in a sucrose or glycerol buffer to which bromophenol blue can be added. This causes the DNA to sink into the well
  • DNA runs from black to red electrode (red is +ve)
  • Gel must have Gel red CAST INTO THE GEL OR POST STAINED AFTER
  • Gels should be 2% (w/v) and be 0.5Ml thick. This will be about 100mL. It can be remelted later.

DNA Extraction for today

Doing all the loris and tawny's and the two other samples picked up today.

  • 7 x loris
  • 1SWWBD
  • 2SWWBD
  • R5#9
  • R5#51
  • R5#52
  • R5#100
  • Pink Gold

And the brown doves

  • RS#93
  • RS#94

Tawnys

  • ethel
  • banjo

I am mixing up a 12x mix of solution 1a and 1b for the EDNA kit. 64ul+16ul is 80ul per tube. Times 12 is 768 of 1a and 192 of 1b.

Notes

Tawnys are fine to DNA sex test (check your bookmarks for the link). Also, the diamond doves may have a problem, as only the males will show up. Maybe in future use a house keeping gene or design some primers around the sequence and get it sanger sequenced.

Results

DNA extraction seems to have worked fine, added the solution 2 and put them in the fridge.