Team:Amsterdam/project/protocols/
From 2012.igem.org
(Difference between revisions)
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'''Day 1''' | '''Day 1''' | ||
- | *1. End of the day: Prepare and incubate (37C) 6x 3ml pSB1AT3+LacH containing bacterial | + | *1. End of the day: Prepare and incubate (37C) 6x 3ml pSB1AT3+LacH containing bacterial cultures (with appropriate antibiotic) |
- | cultures (with appropriate antibiotic) | + | *2. End of the day: Prepare and incubate (37C) 6x 3ml pSB1AT3+pBAD containing bacterial cultures (with appropriate antibiotic) |
- | *2. End of the day: Prepare and incubate (37C) 6x 3ml pSB1AT3+pBAD containing bacterial | + | |
- | cultures (with appropriate antibiotic) | + | |
'''Day 2''' | '''Day 2''' | ||
*1. Start of the day: To 3 cultures from Day 1, Step 1 add to each 15 ul of 1M IPTG solution | *1. Start of the day: To 3 cultures from Day 1, Step 1 add to each 15 ul of 1M IPTG solution | ||
- | *2. Start of the day: To 3 cultures from Day 1, Step 2 add to each 120 ul of 100% (1g/ml) | + | *2. Start of the day: To 3 cultures from Day 1, Step 2 add to each 120 ul of 100% (1g/ml) Arabinose solution |
- | Arabinose solution | + | |
*3. Keep all cultures incubated at 37C | *3. Keep all cultures incubated at 37C | ||
'''Day 3''' | '''Day 3''' | ||
- | *1. Start of day: From 1 of the cultures from Day 2, Step 1 take 2x a 1.65ml sample (1 for | + | *1. Start of day: From 1 of the cultures from Day 2, Step 1 take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (IPTG+ 1 day exposure) |
- | miniprep 1 for western blot), discard rest of culture (IPTG+ 1 day exposure) | + | *2. Start of day: From 1 of the cultures from Day 2, Step 2 take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (IPTG- 1 day negative control) |
- | *2. Start of day: From 1 of the cultures from Day 2, Step 2 take 2x a 1.65ml sample (1 for | + | *3. Start of day: From 1 of the cultures from Day 1, Step 1 (and not used in Day 2, Step 1) take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (ARA+ 1 day exposure) |
- | miniprep 1 for western blot), discard rest of culture (IPTG- 1 day negative control) | + | *4. Start of day: From 1 of the cultures from Day 1, Step 2 (and not used in Day 2, Step 2) take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (ARA- 1 day negative control) |
- | *3. Start of day: From 1 of the cultures from Day 1, Step 1 (and not used in Day 2, Step 1) take | + | *5. Start of day: To the 2 remaining cultures of Day 2, Step 2 each add 120 ul of 100% (1g/ml) Arabinose solution |
- | 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (ARA+ 1 day | + | |
- | exposure) | + | |
- | *4. Start of day: From 1 of the cultures from Day 1, Step 2 (and not used in Day 2, Step 2) take | + | |
- | 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (ARA- 1 day | + | |
- | negative control) | + | |
- | *5. Start of day: To the 2 remaining cultures of Day 2, Step 2 each add 120 ul of 100% (1g/ml) | + | |
- | Arabinose solution | + | |
*6. Keep all (remaining) cultures incubated at 37C | *6. Keep all (remaining) cultures incubated at 37C | ||
'''Day 4''' | '''Day 4''' | ||
- | *1. Start of day: From 1 of the (remaining) cultures from Day 2, Step 1 take 2x a 1.65ml sample | + | *1. Start of day: From 1 of the (remaining) cultures from Day 2, Step 1 take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (IPTG+ 2days exposure) |
- | (1 for miniprep 1 for western blot), discard rest of culture (IPTG+ 2days exposure) | + | *2. Start of day: From 1 of the (remaining) cultures from Day 2, Step 2 take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (IPTG- 2 days negative control) |
- | *2. Start of day: From 1 of the (remaining) cultures from Day 2, Step 2 take 2x a 1.65ml sample | + | *3. Start of day: From 1 of the (remaining) cultures from Day 1, Step 1 (and not used in Day 2, Step 1) take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (ARA+ 2 days exposure) |
- | (1 for miniprep 1 for western blot), discard rest of culture (IPTG- 2 days negative control) | + | *4. Start of day: From 1 of the (remaining) cultures from Day 1, Step 2 (and not used in Day 2, Step 2) take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (ARA- 2 days negative control) |
- | *3. Start of day: From 1 of the (remaining) cultures from Day 1, Step 1 (and not used in Day 2, | + | *5. Start of day: To the 1 remaining culture of Day 2, Step 2 add 120 ul of 100% (1g/ml) Arabinose solution |
- | Step 1) take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture | + | |
- | (ARA+ 2 days exposure) | + | |
- | *4. Start of day: From 1 of the (remaining) cultures from Day 1, Step 2 (and not used in Day 2, | + | |
- | Step 2) take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture | + | |
- | (ARA- 2 days negative control) | + | |
- | *5. Start of day: To the 1 remaining culture of Day 2, Step 2 add 120 ul of 100% (1g/ml) | + | |
- | Arabinose solution | + | |
*6. Keep all (remaining) cultures incubated at 37C | *6. Keep all (remaining) cultures incubated at 37C | ||
'''Day 5''' | '''Day 5''' | ||
- | *1. Start of day: From the 1 remaining culture from Day 2, Step 1 take 2x a 1.65ml sample (1 for | + | *1. Start of day: From the 1 remaining culture from Day 2, Step 1 take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (IPTG+ 3 days exposure) |
- | miniprep 1 for western blot), discard rest of culture (IPTG+ 3 days exposure) | + | *2. Start of day: From the 1 remaining culture from Day 2, Step 2 take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (IPTG- 3 days negative control) |
- | *2. Start of day: From the 1 remaining culture from Day 2, Step 2 take 2x a 1.65ml sample (1 for | + | *3. Start of day: From the 1 remaining culture from Day 1, Step 1 (and not used in Day 2, Step 1) take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (ARA+ 3 days exposure) |
- | miniprep 1 for western blot), discard rest of culture (IPTG- 3 days negative control) | + | *4. Start of day: From the 1 remaining culture from Day 1, Step 2 (and not used in Day 2, Step 2) take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (ARA- 3 days negative control) |
- | *3. Start of day: From the 1 remaining culture from Day 1, Step 1 (and not used in Day 2, Step 1) | + | |
- | take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (ARA+ 3 | + | |
- | days exposure) | + | |
- | *4. Start of day: From the 1 remaining culture from Day 1, Step 2 (and not used in Day 2, Step 2) | + | |
- | take 2x a 1.65ml sample (1 for miniprep 1 for western blot), discard rest of culture (ARA- 3 | + | |
- | days negative control) | + | |
</div> | </div> |
Revision as of 19:18, 23 September 2012