"
Team:University College London/Week14YanikaExp1
From 2012.igem.org
(Difference between revisions)
YanikaBorg (Talk | contribs) (Created page with "For Wnu cell line which has native secreted nuclease activity: 1. Prepare 11-16 plates (10ml LBAgar+10ul AMP +10 ul 1M IPTG). IPTG induces the lac promoter which in turn activa...") |
YanikaBorg (Talk | contribs) |
||
| Line 2: | Line 2: | ||
| - | 1. Prepare 11-16 plates (10ml | + | 1. Prepare 11-16 plates (10ml LB-Agar + 10ul AMP +10ul 1M IPTG). IPTG induces the lac promoter, which in turn activates the transcription of nuclease |
2. Streak cells onto all plates at the same time | 2. Streak cells onto all plates at the same time | ||
| Line 10: | Line 10: | ||
4. Apply hydrochloric acid (HCL) to the first plate before putting them in the incubator (set time as zero) | 4. Apply hydrochloric acid (HCL) to the first plate before putting them in the incubator (set time as zero) | ||
| - | 5. Take a second reading after four hours, followed by six readings every 3 hours, and a final three readings every two hours | + | 5. Take a second reading after four hours, followed by six readings every 3 hours, and a final three readings every two hours |
6. When the reading is taken, observe the following: | 6. When the reading is taken, observe the following: | ||
Revision as of 22:23, 26 September 2012
For Wnu cell line which has native secreted nuclease activity:
1. Prepare 11-16 plates (10ml LB-Agar + 10ul AMP +10ul 1M IPTG). IPTG induces the lac promoter, which in turn activates the transcription of nuclease
2. Streak cells onto all plates at the same time
3. Incubate at 37°C
4. Apply hydrochloric acid (HCL) to the first plate before putting them in the incubator (set time as zero)
5. Take a second reading after four hours, followed by six readings every 3 hours, and a final three readings every two hours
6. When the reading is taken, observe the following:
a) Diameter of the colony (once the diameter of the colony is measured, pick the colony and put it to grow in LB for nine hours)
b) Diameter of the halo that is achieved once HCL is applied
c) OD from a)
d) Estimate of the depth of the colony on the agar plate
