Team:UNITN-Trento/Notebook/Labposts/06/38

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(Created page with "{ "date" : "2013-06-19", "author" : "gabriele-emil-bruno", "title" : "pSB1C3 (quite) an apocalipse and SAMsynthetase the (quite) revenge<html><br/></html>''linearization of pS...")

Revision as of 07:43, 9 September 2013

{ "date" : "2013-06-19", "author" : "gabriele-emil-bruno", "title" : "pSB1C3 (quite) an apocalipse and SAMsynthetase the (quite) revenge
linearization of pSB1C3 ad inocula of SAMsynthetase", "content" : "Today we (GG, ET, BA) tried to linearize pSB1C3 (with poor results), with this protocol, using the circular DNA samples of Tuesday (18-06): Emil 117.3ng/microl, Gabriele 184.1ng/microl.

Then we prepared a gel (1% GellyPhor gel) and checked for the products.

Gel electrophoresis of the products
Loaded samplesResult
1kb ladder-
empty-
B1OK
B2OK
B3OK
E1NO
E2NO
E3NO
G1NO
G2OK
G3OK
empty-
1kb ladder-
Unfortunately, some of the PCR reactions did not succeed!

Then we (GG, ET) have proceeded with the inoculation of pSB1C3+SAMsynthetase (1:1, 1:2, 1:4) and of part R0010 (pLac). Unfortunately there were a few colonies also in the control.
Plate pSB1C3+R0010
Plate pSB1C3+SAMsynthetase ctrl
Plate pSB1C3+SAMsynthetase 1:1
Plate pSB1C3+SAMsynthetase 1:2
Plate pSB1C3+SAMsynthetase 1:4
The inocula were performed in plastic culture tubes (15ml) instead of the glass ones, because we do not know how to sterilize the latter. The tubes were filled with 5ml of LB with Cloramphenycol (1µl of 34mg/ml Cloramphenycol stock solution for 1ml of LB).

In the afternoon Gabriele performed again the same PCR protocol to linearize pSB1C3. He loaded the E1-3 samples again, as double-check.
Gel electrophoresis of the products (Gabriele)
Loaded samplesResult
G1ANO
G2AOK
G3AOK
G4AOK
1kb ladder-
E1NO
E2NO
E3NO
", "tags" : "SAMsynthetase-pSB1C3" }

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