Team:TU Delft/6 July 2010 content

From 2010.igem.org

(Difference between revisions)
(Lab work)
(Lab work)
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<b> Characterization of Anderson RBS sequences </b>
<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.
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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 [http://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 22:58, 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

Pieter by the BBQ
We are ready for the soccer game
the Netherlands won the semi-final!