Template:Team:TU Eindhoven/Protocols HTML

Protocols

For the labwork various protocols were created. These are applied during the work in the Biolab.

General Protocols

• Preparation of general necessities
• Digestion
• Ligation
• PCR Amplification
• Small Culturing
• Transformation into NovaBlue
• Colony PCR

Gibson Assembly

Gibson Assembly is a one-pot assembly method which requires multiple dsDNA fragments as well as a linearized vector. The dsDNA fragments may be generated through PCR or be directly ordered in the form of gBlocks. The vector can either be linearized through PCR or through restriction enzyme digestion. In our case, we ordered gBlocks to obtain the dsDNA fragments and we generated the linear vector through PCR. Therefore, we use gBlocks and vector linearization in our protocols. Another important protocol for Gibson Assembly which is not listed below is Colony PCR.

• Vector Linearization
• Gibson Assembly

Protein Expression

• Double transformation