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Team:TU Delft/6 July 2010 content
From 2010.igem.org
| Line 6: | Line 6: | ||
We were so excited; we immediately solved the DNA in water and performed a [[Team:TU_Delft/protocols/transformation|transformation]] with 100 ng of the DNA and plated on the LB agar containing the appropriate antibiotic. | We were so excited; we immediately solved the DNA in water and performed a [[Team:TU_Delft/protocols/transformation|transformation]] with 100 ng of the DNA and plated on the LB agar containing the appropriate antibiotic. | ||
| - | + | * AlkB2 (Ampicillin) | |
| - | + | * rubA3 (Ampicillin) | |
| - | + | * rubA4 (Ampicillin) | |
| - | + | * rubB (Kanamycin) | |
| - | + | * ladA (Ampicillin) | |
| - | + | * ADH (Ampicillin) | |
| - | + | * ALDH (Kanamycin) | |
| - | + | * bbc1 (Ampicillin) | |
| - | + | * AlnA (Ampicillin) | |
| - | + | * OprG (Ampicillin) | |
| - | + | * PalkS1-2 (Ampicillin) | |
| - | + | * PalkB (Ampicillin) | |
| - | + | * P(CaiF) (Ampenicillin) | |
| - | + | * AlkS (Kanamycin) | |
| - | + | * PhPFD-α (Ampicillin) | |
| - | + | * PhPFD-β (Ampicillin) | |
<h4>Emulsifier</h4> | <h4>Emulsifier</h4> | ||
| Line 79: | Line 79: | ||
<h4>Characterization of Anderson RBS sequences</h4> | <h4>Characterization of Anderson RBS sequences</h4> | ||
| - | Yesterday's purified PCR product of | + | Yesterday's purified PCR product of 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 [https://2010.igem.org/Team:TU_Delft/protocols/ligation ligation] yielded the 5 different RBS sequences now containing the GFP reporter regions. |
==Kampioenen!!!!== | ==Kampioenen!!!!== | ||
Revision as of 18:40, 17 July 2010
Contents |
Lab work
Ordered DNA stocks
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.
- AlkB2 (Ampicillin)
- rubA3 (Ampicillin)
- rubA4 (Ampicillin)
- rubB (Kanamycin)
- ladA (Ampicillin)
- ADH (Ampicillin)
- ALDH (Kanamycin)
- bbc1 (Ampicillin)
- AlnA (Ampicillin)
- OprG (Ampicillin)
- PalkS1-2 (Ampicillin)
- PalkB (Ampicillin)
- P(CaiF) (Ampenicillin)
- AlkS (Kanamycin)
- PhPFD-α (Ampicillin)
- PhPFD-β (Ampicillin)
Emulsifier
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 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
