Difference between revisions of "Preparing an Adenoviral shRNA Clone"
From Bridges Lab Protocols
Davebridges (Talk | contribs) (Created page with '==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<spa...') |
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*<span style="color: #00FFFF">Spacer</span> | *<span style="color: #00FFFF">Spacer</span> | ||
*<span style="color:#FFD700">Loop</span> | *<span style="color:#FFD700">Loop</span> | ||
| + | *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). | 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 | Order primers through IDT-DNA | ||
Revision as of 18:21, 23 August 2010
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
