Designing siRNA for Knockdown

• 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