Transfection of siRNA oligos to peritoneal macrophages

Use DeliverX Transfection from Panomics

Modified protocol below

Step 1:  Make complexes

Note:  Do not scale up more than 3x

Thaw siRNA and DeliverX reagent

Dilute siRNA into buffer 1

10 uM siRNA  2.17 ul

Buffer-1    47.8 ul

Sonicate DeliverX Reagent maximum output for 3-5 min

Dilute into buffer 2

DeliverX Reagent  16 ul

Buffer 2  34ul

Sonicate again for 3-5 min

Mix the diluted siRNA and the diluted DeliverX

Vortex

Incubate at 37 deg C for 20 min

Use immediately or store at -20 deg C for up to 6 months (<10 freeze/thaw)

Mix equal volumes buffer 1+ buffer 2

Add 300 ul mixed buffers to concentrated complexes (total 400 ul)

300

Step 2:  Transfect cells

Incubate 48-72 hours