User:Stuart McKellar/Notebook/Bird Sex Testing/2012/12/03
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Visualising lori's p2/p8 on 1% gelAim: differentiate between male and female loris using PCR product of p2/p8 primers and running on 1% gel. Method: Made up a 1% gel using the life technologies SYBR safe green dye. No dye was added, as the soln was already a 1:1 dilution of dye in TBE buffer. It was easy to make up and dissolved the agar readily. Results: Gels can be seen here: [1] It looks as though the samples A and B are male and that D is female, with the rest being ambiguous.
60°CAim - try to see if results are better at 60°C method: Follwing exact same protocol as here: [2] Master mix for all 8 samples plus -ve control(10 units of MM):
Tubes labelled as follows:
PCR the same except annealing temp is 60°C (user 77 file 81). results to be determined via agar gel E.Phoresis 55CSame as above, samples AT,BT..etc PCR anneal at 55°C |