on Wednesday, February 14th, 2018 1:14 | by Ottavia Palazzo
Map of the phd-ds-Red-attP plasmid used for cloning: 1 desired construct is aiming to target all dFoxP isoforms with one homolgy domain starting at exon 1 to exon 3, the second homology domain starting at exon 3 to exon 6. The second construct is aimed to target just dFoxP IsoB, with both the homology domain at the end of the protein. This plasmid is aimed to knock down the FoxP gene without the Gal-4 knock-in.
PCR product of the homology domains 1 and 2 for FoxP (upper part of the image, left and right), and homology domains 1 and 2 for FoxP-isoformB (lower part of the picture, left and right): amplification of the desired homology domains via PCR (5 replicates for each construct in order to test different annealing temperatures). This products are going to be used for the subsequent cloning in the plasmid.