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Preparing an Adenoviral shRNA Clone

255 bytes added, 19:47, 13 December 2010
→‎Choosing a shRNA Sequence: updated XbaI compatible overhang
__NOTOC__
[[Category:shRNA]][[Category:Adenovirus]][[Category:Transduction]][[Category:Knockdown]][[Category:Molecular Biology]][[Category:Cloning]]
==Choosing a shRNA Sequence==
Use oligoengine to pick a sequence, for example
<span style="color:#FF0000">GATCCTAG</span><span style="color:#00FFFF">CCC</span>GAATACCGCAATGCCTTAG<span style="color:#FFD700">TTCAAGAGA</span>CTAAGGCATTGCGGTATTC<span style="color:#00FFFF">TTTTTA</span>
Where:
==Cloning Targetting Construct==
*see [http://www.oligoengine.com/products/psuper/documentation/protocols/pSUPER_protocol.pdfpSuper] and [http://www.coloncancer.org/adeasy/He,%20AdEasy%20-%20Nature%20Protocols,%202007.pdf AdEasy] protocols
===Digestion of pAdtrack-H1 Promoter (or pSuper)===
#If using a BglII/HindII do a sequential digestion