Team:TU Delft/22 July 2010 content

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Lab work

Alkane degradation

There were some colonies on Tuesday's plates! We had left the plates @ 37°C yesterday after having seen that there were no colonies. When checking this morning on all plates (except the negative control) there were a few colonies! (2-50 colonies). Chances are it's not what we're looking for, but maybe they are good transformants... to check we will do a colony PCR.

1% Agarose gel of colony PCR. Gel runned at 100V for 1 hour. Of all samples 5 μL + 1 μL loadingbuffer was loaded. 5 μL was loaded of marker.
# Description Expected length (bp) Primers Status
1 SmartLadder n/a n/a n/a
2 Transformant #1 of ligation mix 007T 1616 G00100 + G00101
3 Transformant #2 of ligation mix 007T 1616 G00100 + G00101
4 Transformant #3 of ligation mix 007T 1616 G00100 + G00101
5 Transformant #1 of ligation mix 008T 551 G00100 + G00101
6 Transformant #2 of ligation mix 008T 551 G00100 + G00101
7 Transformant #1 of ligation mix 009T 551 G00100 + G00101
8 Transformant #1 of ligation mix 009T 560 G00100 + G00101
9 Transformant #1 of ligation mix 009T 560 G00100 + G00101
10 Transformant #1 of ligation mix 010T 1657 G00100 + G00101
11 Transformant #1 of ligation mix 010T 1657 G00100 + G00101
12 Transformant #1 of ligation mix 010T 1657 G00100 + G00101
13 Transformant #1 of ligation mix 017T 1130 G00100 + G00101
14 Transformant #1 of ligation mix 017T 1130 G00100 + G00101
15 Transformant #1 of ligation mix 017T 1130 G00100 + G00101
16 Transformant #1 of ligation mix 018T 1874 G00100 + G00101
17 Transformant #1 of Red colony 1360 G00100 + G00101

A number of colonies look promising! To check if they really are the BioBricks we want, tomorrow we will do a plasmid isolation with the cultures of lane 2, 4, 6, 7, 9 and 14. We will cut the isolated plasmids with various restriction enzymes and analyze the digestion products on gel.