Team:Groningen/Notebook/tasA Digestion K23 t44
Blue Bio Energy
tasA Digestion K823023 (t44)
The miniprepped samples were digested to check for the right insert.
Our goal is to overexpress TasA by adding a second copy of the tasA gene under the control of a salt inducible promoter. This overexpression will lead to a stronger biofilm when the biofilm is exposed to salt water.
tasA was inserted successfully into pSB1C3.
Restriction
00:00, 10 July 2015 - 00:00, 10 July 2015
The miniprepped samples were digested to check for insert. It was seen that indeed tasA was inserted successfully into pSB1C3. The insert was stored as T2.
00:00, 16 July 2015 - 00:00, 16 July 2015
The salt promoter, tasA (T2) and the backbone were digested. For the digest the following combinations were made.
#
DNA
Restriction enzymes
1
Psalt
EcoRI + SpeI
2
tasA (T2, 10-7-15)
XbaI + PstI
3
BBa_K823023
EcoRI + PstI
Digestion.
#
Component
Amount
1
DNA (Psalt)
6 µL
EcoRI
1 µL
SpeI
1 µL
10x buffer (2.1)
2 µL
\( \mathrm{H_2O}\)
10 µL
2
DNA (T2)
3 µL
XbaI
1 µL
PstI
1 µL
10x buffer (2.1)
2 µL
\( \mathrm{H_2O}\)
13 µL
3
DNA (K823023)
6 µL
EcoRI
1 µL
PstI
1 µL
10x buffer (2.1)
2 µL
\( \mathrm{H_2O}\)
10 µL
Components samples.
The samples were incubated for one hour at 37 °C.