Team:Warsaw/Calendar-Main/1 October 2009
From 2009.igem.org
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Revision as of 18:20, 12 October 2009
Contents |
Isolation of BBa_J63010 from 2009 Kit
Monika
Task:
- Isolate BioBrick BBa_J63010 from 2009 Kit Plate 1
Methods:
- Alkaline lysis with A&A Biotechnology Kit, procedure described here
- Control digestion of BBa_J63010
First reaction mixture composition:
5μl plasmid solution 0,3μl EcoRI (Fermentas) 0,3μl PstI (Fermentas) 2μl Buffer Orange (Fermentas) 12,4 μl MQ water
Second reaction mixture composition:
5μl plasmid solution 0,5μl PvuII (Fermentas) 2μl Buffer Green (Fermentas) 12,3 μl MQ water
Digestion in 37°C for 2,5h
Bistable switch testing
Monika
Tasks:
- Planting GFP expressing clones on LB medium with tetracycline
Cloning of the cro-box into the pSB1A3 plasmid
Kamil
Tasks:
- Plasmid isolation
- Control digest
Methods:
- The plasmids were isolated from 3μl of overnight liquid cultures using the Plasmid Mini kit (A&A Biotechnology).
- The digest mix was prepared as follows:
20μl purified plasmid
3μl Orange Buffer (Fermentas)
1μl PstI Enzyme (Fermentas)
1μl EcoRI Enzyme (Fermentas)
5μl H2O- The digest was carried out in 37°C for 4h.
- The results were visualized on 2% agarose gel.
Results:
-
(Yes, a filler :) )
- Gel (from left):
- GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
- Sample 1
- Sample 2
- Sample 3
- Sample 4
- Sample 5
- Reference sample
- Sample 6
- Sample 7
- Sample 8
- Sample 9
Conclusions:
- It all looks promising, samples 1, 2, 3, and 7 were sent to be sequenced.
[UPDATE:] The nucleotides in sample 7 read: "tacctctggcggtgata" (prefix and suffix omitted), which means that we have a proper cro-box!.
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