Team:TUDelft/4 September 2009

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4 September 2009

Tim Weenink

I transformed all four batches of competent cells with !SIII ligation mix. Plates (Cam) were left on the bench in a plastic bag over the weekend. All the time constants in the electroporation were 4.4 or 4.5 (which is good). Also two plates without selection marker were streaked to check if the competent cells were viable at all.

Saeed

I made minipreps form 6 overnight cultures of !P in order to check the assembly of !P. These are called !P1 to !P6.


Part [DNA] ng/uL
 !P1 1001.1
 !P2 92.3
 !P3 98.0
 !P4 102.2
 !P5 89.1
 !P6 86.1

The isolated plasmide were restricted with E and P in order to check fragment size. !P1, !P3 and !P5 are also cut upstream.

Saeed040909.jpg


lane Part Expected Plasmid Size Status
1 [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder]
2  !P1 833 ok
3  !P2 833 ok
4  !P3 833 ok
5  !P4 833 ok
6  !P5 833 ok
7  !P6 833 ok
8 [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder]
9  !P1 upstream 833 ok
10  !P3 upstream 833 ok
11  !P5 upstream 833 ok
12 [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder]

The picture is not realy clear but the 833 bp fragment was visible.

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