Fusion-based cloning in SnapGene tutorial

From BITS wiki
Jump to: navigation, search
Go to parent SnapGene

SnapGene has a list of In-Fusion ready linearized vectors from Clontech on its website.

The file containing the vector that we are going to use, pAcGFP1-N In-Fusion® Ready, is available in the folder /Documents/SnapGene . Load the sequence in SnapGene.


Open the cloning editor


Prepare the vector sequence

The cloning editor opens on the Vector tab where the vector that was loaded when you opened the cloning editor (pAcGFP1-N In-Fusion® Ready) is opened automatically. Since this is an In-Fusion vector it is already linearized. If this was not the case you could linearize the vector by PCR or restriction digest. SG57.png


Prepare the insert

We are going to clone Rab5 into this vector thereby created a N-terminal fusion of Rab5 to AcGFP1.

You do need primers to amplify Rab5 from the mRFP1Rab5 vector creating overlaps with the In-Fusion vector. SnapGene can design these primers automatically.


Perform the cloning


Show a graphical overview of the cloning procedure


Obtain the sequences of the primers

See the In-Fusion cloning video tutorial for further details on how SnapGene can simulate Clontech's In-Fusion Cloning while helping you to visualize, control, and document the process.