"
Team:Warsaw/Calendar-Stage1/5 July 2010
From 2010.igem.org
Confirmation of biobricks obtained from Distribution [Kasia, Jarek]
1.Finish alkaline lysis no3.
2.Run 5ul of undigested plasmids on the agarose gel.
[tu powinien byc zalacznik w postaci obrazka]
Correct amount of plasmids, expected size.
Cloning of GFP-terminator behind RBSes - approach 1 -Anderson's RBSes [Ania1, Ania2, Ania3, Jarek; Kasia, Michal]
1. Rescue samples from alkaline lysis no.2 by adding RNAse.
2. SpeI/PstI digest of samples 2-11 (RBSes:). time:3h
3. XbaI/PstI digest of sample 1 (GFP+terminator). time:3h
4. Run GFP+terminator on agarose gel [tu powinien byc zalacznik w postaci obrazka] Small amount of plasmid.
5. Gel extraction of GFP+terminator.
6. Thermal inactivation of samples 2-6 (Anderson's RBSes) after SpeI/PstI digest.
7. Ligation of digested samples 2-6 (Anderson's RBSes) with gel-extracted GFP-terminator
8. Transformation with the ligation mixtures.
Cloning of GFP-terminator behind RBSes - approach 2[Ania1, Ania2, Ania3, Jarek; Kasia
]
1. Set up overnight GFPterminator digest with XbaI/PstI. [16ul DNA from alkaline lysis no3, 1ul XbaI, 1ulPstI, 4ul BamHI buffer, water up to 40ul.]
Prepare glycerol stocks from samples 1-12 [Ania1, Ania2, Ania3, Jarek; Kasia
1. Set up 2ul liquid culture of each biobrick.
