Preparing an Adenoviral shRNA Clone: Difference between revisions
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__NOTOC__ | __NOTOC__ | ||
[[Category:shRNA]][[Category:Adenovirus]][[Category:Transduction]][[Category:Knockdown]][[Category:Molecular Biology]][[Category:Cloning]] | |||
==Choosing a shRNA Sequence== | ==Choosing a shRNA Sequence== | ||
Use oligoengine to pick a sequence, for example | Use oligoengine to pick a sequence, for example | ||
<span style="color:#FF0000"> | <span style="color:#FF0000">CTAG</span><span style="color:#00FFFF">CCC</span>GAATACCGCAATGCCTTAG<span style="color:#FFD700">TTCAAGAGA</span>CTAAGGCATTGCGGTATTC<span style="color:#00FFFF">TTTTTA</span> | ||
Where: | Where: | ||
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==Cloning Targetting Construct== | ==Cloning Targetting Construct== | ||
*see http://www.oligoengine.com/products/psuper/documentation/protocols/pSUPER_protocol.pdf | *see [http://www.oligoengine.com/products/psuper/documentation/protocols/pSUPER_protocol.pdf pSuper] and [http://www.coloncancer.org/adeasy/He,%20AdEasy%20-%20Nature%20Protocols,%202007.pdf AdEasy] protocols | ||
===Digestion of pAdtrack-H1 Promoter (or pSuper)=== | ===Digestion of pAdtrack-H1 Promoter (or pSuper)=== | ||
#If using a BglII/HindII do a sequential digestion | #If using a BglII/HindII do a sequential digestion | ||