more info about pHD-ScarlessDsRed

[Jihyun Irizarry]

Hello,

I am trying to tag a gene of my interest using CRISPR/Cas9 system.

To insert fluorescent protein at the C-terminus, I'd like to use pHD-ScarlessDsRed donor construct.

However, I haven't found a good protocol how to modify this donor construct to clone in homology arms.

Does anyone have any suggestion for ref. or protocol?

Thank you!

Jihyun

[Marlen Schmidt]

You might try Red/ET recombination (http://www.genebridges.com/recombineering), which allows for flexible and nucleotide precise DNA modification at any position, espacily for modification of large and complex constructs. Insertion of (large) homology arms are possible as well.