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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
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.
Transformation with the ligation mixtures.
Summarised by Milena.
