A1CF Human siRNA

siRNA refers to small interfering or short interfering RNA. siRNA Gene Silencers are pools of three target specific 19-25 nucleotide-long double stranded RNA molecules with 2-nt 3' overhangs on each end. The siRNA Gene Silencers are provided as 3 vials of 5nmol (15nmol) in lyophilized form. Each vial contains slightly different sequences to ensure full knockout of the gene. The duplexes can be transfected individually or pooled together to achieve knockdown of the target gene, which is most commonly assessed by qPCR or western blot.

Each siRNA oligo is purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex, and to ensure maximum lot-to-lot consistency. Each lot is monitored base by base through trityl analysis to ensure appropriate coupling efficiency.

IFU: Transfection with 10nM-100nM siRNA, 48 to 72 hours prior to cell lysis. Before re-suspending, briefly centrifuge the tube to ensure the lyophilized siRNA is at the bottom of the tube. Resuspend the siRNA oligos to an appropriate concentration with DEPC water. For example, resuspend one tube of 5nmol siRNA oligo in 250ul of DEPC water to get a final concentration of 20uM. Each vial is suitable for 250 transfections in 24 well-plate (20pmol for each well).