Team:Groningen/Template/MODULE/Notebook/secretion/week9
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September 29 - October 5
goal: Obtaining pLASs with 3-a assembly
Extra amounts of dspB, dspB+HIS, aiiA, aiiA+HIS,aiiA+HIS, pLAS, RBS, double terminator, pSB2k3, pSB1C3 and pSB1A3 were miniprepped to be used for the assembly
1 ug of aiiA, dspB, dspB+HIS and aiiA+HIS with XbaI and PstI and 1 ug of RBS was cut with EcoRI and SpeI
These were eventually ligated for 3 hours into pSB2K3 and transformed into E. coli NEB cells:
Name assembly | Upstream part | Downstream part | Backbone |
---|---|---|---|
A1’ | RBS | aiiA | pSB2K3 |
A2’ | RBS | dspB | pSB2K3 |
A3’ | RBS | dspB+HIS | pSB2K3 |
A4’ | RBS | aiiA+HIS | pSB2K3 |