2shRNAtrc

2shRNA Oligo Designer for pLKO, pQCtrc, and others using 21-mer shRNA target

Note: designer for pLentilox and other 19-mer vector is running at http://en.dogeno.us/2shRNA

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designer for pLKO.1, using AgeI/EcoRI insertion sites

target symbol:

• siRNA sequence
  
• 21 nt length check
  
• G as the first nt, not required in pLKO
  
• loop region
  
• 5 end enzyme 5-
  |
  -3 (leave blank if it is blunt; default ligated to AgeI cut 5-A | CCGGT-3)
• 3 end enzyme site 5-
  |
  -3 (sticky end represents as 5-G | AATTC-3 for EcoRI)

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designer for pLKO.3G, using EcoRI/PacI insertion sites

target symbol:

• siRNA sequence
  
• 21 nt length check
  
• G as the first nt, not required in pLKO
  
• loop region
  
• 5 end enzyme 5-
  |
  -3 (leave blank if it is blunt; default ligated to EcoRI cut 5-G | AATTC-3)
• 3 end enzyme site 5-
  |
  -3 (sticky end represents as 5-TTAAT | TAA-3 for PacI)