User:Sim Huay Ping/Notebook/CBE/08/150/2008/12/04: Difference between revisions
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==Bacteria and AMP storage== | ==Bacteria and AMP storage== | ||
1. From the streak plates, extract 1 colony of E.Coli using the inoculating loop and dip the inoculating loop into 4ml of MHB (for bacteria) and Sabourand Dextrose Broth (for yeast). | 1. From the streak plates, extract 1 colony of E.Coli using the inoculating loop and | ||
dip the inoculating loop into 4ml of MHB (for bacteria) and Sabourand Dextrose | |||
Broth (for yeast). | |||
2. Do the same for Staphylococcus, Pseudomonas Aeruginosa and Candida. | 2. Do the same for Staphylococcus, Pseudomonas Aeruginosa and Candida. | ||
3. Put the bacteria tubes in the rotating incubator at 37 degC. | 3. Put the bacteria tubes in the rotating incubator at 37 degC. | ||
Put the yeast tubes in the dry incubator at 28 degC. | |||
Put the yeast tubes in the dry incubator at 28 degC. | |||
4. Collect the inoculums after 5-6 hours. | 4. Collect the inoculums after 5-6 hours. | ||
5. Extract 1000ul of inoculum into the vials and centrifuge the vials at 1000rpm for 5 minutes. | |||
5. Extract 1000ul of inoculum into the vials and centrifuge the vials at 1000rpm for | |||
5 minutes. | |||
6. Remove the supernatant and then add in 1000ul of PBS into the vials. | 6. Remove the supernatant and then add in 1000ul of PBS into the vials. | ||
7. Vortex and centrifuge again. | 7. Vortex and centrifuge again. | ||
8. Repeat Steps 6 and 7 for three times. | 8. Repeat Steps 6 and 7 for three times. | ||
9. Vortex after adding the last 1000ul of PBS to ensure continuous mixture. | 9. Vortex after adding the last 1000ul of PBS to ensure continuous mixture. | ||
10. Extract 500ul of the samples into another vial. This makes 2 vials of equal amounts. | 10. Extract 500ul of the samples into another vial. This makes 2 vials of equal amounts. | ||
11. Label “Empty with the bacteria’s name” on the first vial sample and label “bacteria’s name and AMP on the second vial sample. | |||
11. Label “Empty with the bacteria’s name” on the first vial sample and label | |||
“bacteria’s name and AMP on the second vial sample. | |||
12. Incubate for 15 minutes. | 12. Incubate for 15 minutes. | ||
13. Put the vials into the freezer. | 13. Put the vials into the freezer. | ||
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Bacteria and AMP storage1. From the streak plates, extract 1 colony of E.Coli using the inoculating loop and dip the inoculating loop into 4ml of MHB (for bacteria) and Sabourand Dextrose
Broth (for yeast).
2. Do the same for Staphylococcus, Pseudomonas Aeruginosa and Candida. 3. Put the bacteria tubes in the rotating incubator at 37 degC. Put the yeast tubes in the dry incubator at 28 degC. 4. Collect the inoculums after 5-6 hours. 5. Extract 1000ul of inoculum into the vials and centrifuge the vials at 1000rpm for 5 minutes. 6. Remove the supernatant and then add in 1000ul of PBS into the vials. 7. Vortex and centrifuge again. 8. Repeat Steps 6 and 7 for three times. 9. Vortex after adding the last 1000ul of PBS to ensure continuous mixture. 10. Extract 500ul of the samples into another vial. This makes 2 vials of equal amounts. 11. Label “Empty with the bacteria’s name” on the first vial sample and label “bacteria’s name and AMP on the second vial sample. 12. Incubate for 15 minutes. 13. Put the vials into the freezer.
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