Template:Team:Groningen/CONTENT/LOGBOOK/Gibson Assembly pg30
Gibson assembly (pg30)
With a Gibson assembly two pieces of DNA are combined. PGA was ordered in two different parts. These parts are combined with a Gibson assembly.
Make a more negatively charged biofilm.
Gibson assembly failed.
<a class="postscriptum protocol" href="https://2015.igem.org/Team:Groningen/Protocols_and_Protocols/Gibsonassembly">Gibson assembly</a>
12:00, 10 August 2015 - 15:00, 10 August 2015
Gibson assembly of PGA. The following Gibson Assembly master mix was prepared for 5 reactions.
Compound
Amount
5X ISO buffer
20µL
Ts exe
2µL
Phusion
1.25µL
Taq ligase
10µL
H2O
2µL
Reaction master mix for Gibson assembly.
Each reaction 15 ul Gibson assembly mix + 5 ul DNA (2,5µL PGA 1 and 2,5µL PGA 2).
Both products were amplified was also directly used for bacillus transformation according to protocol ( 10 ul was used for transformation)
PCR primers used: Prefix and suffix primers