Designing siRNA for Knockdown
From Bridges Lab Protocols
Revision as of 22:37, 8 February 2012 by Davebridges (Talk | contribs) (added instructions for reordering a siRNA)
- Use Invitrogen's BLOCK-IT RNAi Designer [1]
- Find the refseq RNA sequence for your gene by going to Entrez Gene and entering the gene name and sequence. The RefSeq sequence should start with "NM_"
- Select the appropriate control species and whether to target the coding sequence (normally what we do), or the UTRs
- You will usually get a list of several potential sequences, ordered by location on the RNA molecule. Pick 2-3 sequences, based on the alternative splicing of the RNA. If there is only one listed splice variant on NCBI Gene, pick one towards the N-terminus and one towards the C-terminus.
- Select continue
- Click the "tube" button and disregard the scrambled RNA controls.
- Click "Order Online" button.
- Add your name to Reseacher Name and order.
- Use Promo code P817600 until 30 June 2012.
Reordering a Stealth siRNA
- Go here https://products.invitrogen.com/ivgn/en/US/adirect/invitrogen?cmd=ConfigureRNASingleTube
- Select 3' Addition to dTdT
- Select Duplex
