Preparing an Adenoviral shRNA Clone

Choosing a shRNA Sequence

Use oligoengine to pick a sequence, for example GATCCCCGAATACCGCAATGCCTTAGTTCAAGAGACTAAGGCATTGCGGTATTCTTTTTA

Where:

• XbaI compatible overhang
• Spacer
• Loop
• In black is the target shRNA sequence, both forward and then after the loop as a reverse complement.

For the reverse primer choose the reverse complement, remove the XbaI overhang from the 3' end (the last GATC sequence) and add an AGTC sequence to the 5' end (for HindIII). Order primers through IDT-DNA