Difference between revisions of "SDS Page-Gel for all the bovine proteins"
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Materials: | Materials: | ||
Casein | Casein | ||
Line 20: | Line 21: | ||
control 5FOA | control 5FOA | ||
transformed negative control | transformed negative control | ||
liquid culture | |||
Bovine Beta B (8) - 20ml + 4ml loading dye | |||
Bovine Alpha A (8) - 20ml + 4ml loading dye | |||
Method: | Method: | ||
Get 3 colonies from each plates in 1 centrifuge tube + resuspend into 20ml distilled water | Get 3 colonies from each plates in 1 centrifuge tube + resuspend into 20ml distilled water | ||
For casein touch 1 20-2w microliter pipette tip and rinse with 20 microliter dionised water in 1 casein control tube | For casein touch 1 20-2w microliter pipette tip and rinse with 20 microliter dionised water in 1 casein control tube | ||
Line 29: | Line 37: | ||
To run the gel | To run the gel | ||
Take 20 microliter of sample and 4 microliter loading dye | |||
Take 20 microliter of sample and 4 microliter of loading dye | |||
Boil all the samples for 10 minutes | Boil all the samples for 10 minutes | ||
Total amount of loading (1x) buffer we required 400 ml (10 ml of 10x L. Buffer into 90 ml distilled water) | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Lane''' | |||
| align="center" style="background:#f0f0f0;"|'''1''' | |||
| align="center" style="background:#f0f0f0;"|'''2''' | |||
| align="center" style="background:#f0f0f0;"|'''3''' | |||
| align="center" style="background:#f0f0f0;"|'''4''' | |||
| align="center" style="background:#f0f0f0;"|'''5''' | |||
| align="center" style="background:#f0f0f0;"|'''6''' | |||
| align="center" style="background:#f0f0f0;"|'''7''' | |||
| align="center" style="background:#f0f0f0;"|'''8''' | |||
| align="center" style="background:#f0f0f0;"|'''9''' | |||
| align="center" style="background:#f0f0f0;"|'''10''' | |||
|- | |||
| ||Ladder||5FOA Control||Control (transformed)||Gene (1)||Gene (8)||Gene (3B)||Gene (6B)||Gene (7)||Casein ||Bovine B (liquid culture) | |||
|- | |||
| | |||
|} | |||
Run the gel on 150v for almost 1 hour | |||
Staining and destaining: Coomasie stain solution, destain solution kimwipes (or something else to absorb the coomassie) | |||
Result: | Result: We didn't get result on gel |
Latest revision as of 23:36, 7 December 2014
SDS Page-Gel for all of the bovine proteins
Monday Nov 24th
SDS Page-Gel for all of the bovine proteins
Participants: Wesley, Meenakshi, Nikola, Johan
Materials:
Casein
3B(100 fold diluted)
6B (100 fold diluted)
7 (100 fold diluted)
2B (100 fold diluted)
control 5FOA
transformed negative control
liquid culture
Bovine Beta B (8) - 20ml + 4ml loading dye
Bovine Alpha A (8) - 20ml + 4ml loading dye
Method:
Get 3 colonies from each plates in 1 centrifuge tube + resuspend into 20ml distilled water For casein touch 1 20-2w microliter pipette tip and rinse with 20 microliter dionised water in 1 casein control tube One sample take 1 colony from SFOA (as a negative control) One sample from control yeast transform plate
To run the gel
Take 20 microliter of sample and 4 microliter of loading dye Boil all the samples for 10 minutes
Total amount of loading (1x) buffer we required 400 ml (10 ml of 10x L. Buffer into 90 ml distilled water)
Lane | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 |
Ladder | 5FOA Control | Control (transformed) | Gene (1) | Gene (8) | Gene (3B) | Gene (6B) | Gene (7) | Casein | Bovine B (liquid culture) | |
Run the gel on 150v for almost 1 hour
Staining and destaining: Coomasie stain solution, destain solution kimwipes (or something else to absorb the coomassie)
Result: We didn't get result on gel