User:Stuart McKellar/Notebook/Bird Sex Testing/2012/12/03
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(→Visualising lori's p2/p8 on 1% gel) |
Current revision (03:53, 3 December 2012) (view source) (→Visualising lori's p2/p8 on 1% gel) |
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*B:2SWWBD | *B:2SWWBD | ||
*D:R5#51 | *D:R5#51 | ||
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| + | ==60°C== | ||
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| + | Aim - try to see if results are better at 60°C | ||
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| + | method: | ||
Follwing exact same protocol as here: | Follwing exact same protocol as here: | ||
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*DX:R5#51 | *DX:R5#51 | ||
*EX:R5#52 | *EX:R5#52 | ||
| - | *FX:R5#100 - i htink i missed this one | + | *FX:R5#100 - (i htink i missed this one) |
*GX:Pink Gold | *GX:Pink Gold | ||
*HX:banjo | *HX:banjo | ||
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results to be determined via agar gel E.Phoresis | results to be determined via agar gel E.Phoresis | ||
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| + | ==55C== | ||
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| + | Same as above, samples AT,BT..etc | ||
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| + | PCR anneal at 55°C | ||
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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 | |
