Template:Team:Groningen/CONTENT/LOGBOOK/tasA Ligation t46
Ligation tasA, Psalt in K823023 (t46)
In this experiment the salt promoter is ligated with tasA in the K823023 backbone.
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.
The ligation of tasA , Psalt and K823023 was performed.
<a class="postscriptum protocol" href="https://2015.igem.org/Team:Groningen/Protocols_and_Protocols/Ligation">Ligation</a>
00:00, 16 July 2015 - 00:00, 16 July 2015
The samples 1, 2 and 3 from the digestion step were ligated using the following amounts.
Component
Amount
DNA 1 (Psalt)
5 µL
DNA 2 (T2)
5 µL
DNA 3 (K823023)
2.5 µL
T4 ligase
0.5 µL
10x buffer
2 µL
\( \mathrm{H_2O}\)
5 µL
The sample was ligated at room temperature for 30 minutes.