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Team:Warsaw/Calendar-Stage1/5 July 2010
From 2010.igem.org
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<br>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.] | <br>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.] | ||
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| - | <br><div class="note"> Prepare glycerol stocks</div> | + | <br><div class="note"> Prepare glycerol stocks from samples 1-12</div> |
<br>1. Set up 2ul liquid culture of each biobrick. | <br>1. Set up 2ul liquid culture of each biobrick. | ||
Revision as of 22:05, 9 July 2010
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]
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
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
1. Set up 2ul liquid culture of each biobrick.
