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Team:TU Delft/6 July 2010 content
From 2010.igem.org
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| + | <b> Characterization of Anderson RBS sequences </b> | ||
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| + | Yesterday's purified PCR product of BBa_I13401 was [[digested]] using XbaI and PstI in order to obtain a back-insert. The relevant Anderson RBS Biobricks (J61100, J61101, J61107, J61117 and J61127) were digested using SpeI and PstI. Purification of the digestion products using Roche's PCR Purification Kit and subsequent overnight ligation yielded the 5 different RBS sequences now containing the GFP reporter regions. | ||
===Kampioenen!!!!=== | ===Kampioenen!!!!=== | ||
Revision as of 20:34, 12 July 2010
Lab work
The DNA from Mr Gene has finally arrived, now we're ready to build some biobricks!
We were so excited; we immediately solved the DNA in water and performed a transformation with 100 ng of the DNA and plated on the LB agar containing the appropriate antibiotic.
1. AlkB2 on 100 μg/mL Ampicillin
2. rubA3 on 100 μg/mL Ampicillin
3. rubA4 on 100 μg/mL Ampicillin
4. rubB on 50 μg/mL Kanamycin
5. ladA on 100 μg/mL Ampicillin
6. ADH on 100 μg/mL Ampicillin
7. ALDH on 50 μg/mL Kanamycin
8. bbc1 on 100 μg/mL Ampicillin
9. AlnA on 100 μg/mL Ampicillin
10. OprG on 100 μg/mL Ampicillin
11. PalkS1-2 on 100 μg/mL Ampicillin
12. PalkB on 100 μg/mL Ampicillin
13. P(CaiF) on 100 μg/mL Ampenicillin
14. AlkS on 50 μg/mL Kanamycin
15. PhPFD-α on 100 μg/mL Ampicillin
16. PhPFD-β on 100 μg/mL Ampicillin
Pieter is testing different condition for the emulsifying Assay. To determine the emulsifying activity we set up a calibration curve with SDS. He prepared the following samples:
| Hexane (mL) | Tris Buffer pH 8 (mL) | 10% SDS (mL) | |
| B | 1 | 1.1 | 0 |
| 100 | 1 | 1 | 0.1 |
| 50 | 1 | 1.05 | 0.05 |
| 25 | 1 | 1.075 | 0.025 |
| 10 | 1 | 1.09 | 0.01 |
| 5 | 1 | 1.095 | 0.005 |
Characterization of Anderson RBS sequences
Yesterday's purified PCR product of BBa_I13401 was digested using XbaI and PstI in order to obtain a back-insert. The relevant Anderson RBS Biobricks (J61100, J61101, J61107, J61117 and J61127) were digested using SpeI and PstI. Purification of the digestion products using Roche's PCR Purification Kit and subsequent overnight ligation yielded the 5 different RBS sequences now containing the GFP reporter regions.
Kampioenen!!!!
Today the Dutch team will play the semi-final of the World Cup! Hear us cheer! Stop sound
