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Team:Warsaw/Calendar-Stage1/6 July 2010
From 2010.igem.org
6.07.2010 Tuesday
Preparation of glycerol stocks [Dominik, Ania S., Kasia]
1.Glycerol stocks of following biobricks: J61100, J61101, J61107, J61117, J61127 [Anderson's RBSes], B0030, B0031, B0032, B0033, B0034 [community RBSes], J23100 [synthetic promoter].
2. Alkaline lysis of part of liquid culture used to prepare stocks. Undigested plasmids run on the gel. Results seem to be correct.
Cloning GFP-terminator behind RBSes - approach 2 [Ania P, Ania O., Kasia, Milena]
1. Digest of GFP-terminator in psb1A2 plasmid. [16 μl DNA, 6 μl buforu, 16 μl H2O, 0,5 μl Xba/ Pst]
2. GFP-terminator gel extraction.
3. GFPterminator ligation with B0030, B0031, B0032, B0033, B0034 [community RBSes], [1 μl vector-RBSes, 10 μl < insert-GFPterminator, 2 μl buffer, 7 μl water]
4. Transformation with 20 μl ligation mixture
Cloning GFP-terminator behind RBSes - approach 1 [Ania S., Ania P., Kasia]
05.07.2010 transformation results: contamination with ampicillin susceptible bacteria.
Set up one liquid culture (3,5 ml) from each plate. Cultures inocculated with mix of all clones obtained on the plate.
-> Repeated ligation of digested samples 2-6 (Anderson's RBSes) with gel-extracted GFP-terminator [Michał].
-> Transformation with the ligation mixtures.
Summarised by Milena.
