Team:Warsaw/Calendar-Main/15 October 2009
From 2009.igem.org
Creation of the cro-box testing devices
Kamil
Tasks:
- Reporter device carrying plasmid digestion
- Cro-box carrying plasmids digestion
Methods:
- For the reporter device the digest mix was prepared as follows:
10μl purified plasmid
2μl Tango Buffer (Fermentas)
1μl XbaI Enzyme (Fermentas)
1μl PstI Enzyme (Fermentas)
5μl H2O- For the cro-box the digest mix was prepared as follows (same for both plasmids):
10μl purified plasmid
2μl Tango Buffer (Fermentas)
1μl BcuI Enzyme (Fermentas)
1μl PstI Enzyme (Fermentas)
5μl H2O- The digest was carried out in 37°C for 4h.
- The results were visualized on 1% agarose gel.
- Gel extraction and purification of the appropriate bands was carried out using the Gel-Out kit (A&A Biotechnology)
Mutagenesis of cIKuba
Task: Obtain a functional, termosensitive version of cI
- inactivation ligation (15', 65oC)
- Dpn1 digest (5 h)
- transformation into E. coli (Top10)
April M T W T F S S 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 May M T W T F S S 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 June M T W T F S S 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 July M T W T F S S 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 August M T W T F S S 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 September M T W T F S S 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 October M T W T F S S 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 - For the cro-box the digest mix was prepared as follows (same for both plasmids):