View source for Preparing an Adenoviral shRNA Clone

==Choosing a shRNA Sequence==
Use oligoengine to pick a sequence, for example
<span style="color:#FF0000">GATC</span><span style="color:#00FFFF">CCC</span>GAATACCGCAATGCCTTAG<span style="color:#FFD700">TTCAAGAGA</span>CTAAGGCATTGCGGTATTC<span style="color:#00FFFF">TTTTTA</span>

Where:
*<span style="color: #FF0000">XbaI compatible overhang</span>
*<span style="color: #00FFFF">Spacer</span>
*<span style="color:#FFD700">Loop</span>

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