User:Stuart McKellar/Notebook/Bird Sex Testing/2012/11/13
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| - | == | + | ==gel electrophoresis== |
| - | * | + | * The aim of this lab is to determine the sex of the rainbow lorikeets through agar gel electrophoresis. Gel is 2% agar and stained with GelGreen that has been sitting on my desk room temperature since last test (which was successful). I will run a ladder as well to make sure that the gel is still showing luminescence. |
| + | If the PCR failed, I expect the DNA not to migrate. I am loading 5ul DNA sample with 1ul x 6x DNA running buffer (glycerol + bromophenol blue). | ||
| + | |||
| + | Samples were loaded as follows: | ||
| + | *1SWWBD | ||
| + | *2SWWBD | ||
| + | *R5#9 | ||
| + | *Ladder (300-10KB) | ||
| + | *R5#51 | ||
| + | *R5#52 | ||
| + | *R5#100 | ||
| + | *Pink Gold | ||
| + | |||
| + | Gel ran at 100V (intially started at 300V for a few seconds and then pansied out) and 60mA. Immediate illumination could be seen in the well with the ladder. Gel is running and I can see the ladder but nothing else. Fuck. Looks like I am going back to PCR. After 300bp had reached centre, still no light from samples. Maybe run a single lorikeet sample through PCR and then check on a gel. | ||
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gel electrophoresis
If the PCR failed, I expect the DNA not to migrate. I am loading 5ul DNA sample with 1ul x 6x DNA running buffer (glycerol + bromophenol blue). Samples were loaded as follows:
Gel ran at 100V (intially started at 300V for a few seconds and then pansied out) and 60mA. Immediate illumination could be seen in the well with the ladder. Gel is running and I can see the ladder but nothing else. Fuck. Looks like I am going back to PCR. After 300bp had reached centre, still no light from samples. Maybe run a single lorikeet sample through PCR and then check on a gel. | |
