Infographic: non-viral cell transfection and gene delivery

Protocols

Make yourself over £12,000 worth of electroporation buffer for about £1

Commercially available mammalian electroporation buffer is horrifically expense. We used to use this buffer that is priced at £646 for 25 ml.

Supplier listing: Ingenio Electroporation Solution, 25 ml (MIR 50117), catalogue E7-0520, price £646.00

We now make 500ml of buffer (which would cost £12920 if you purchased 500 ml of the commercial buffer) as detailed below for about £1 and in our hands transfecting fibroblasts and smooth muscle cells it gives better transfection efficiency and similar, if not better viability than the commercial stuff.

Protocol

  1. Get an old, empty (sterile) 500 ml PBS bottle from the tissue culture lab.
  2. Add the reagents in the table below.
    Reagent table for 500 ml electroporation buffer: anhydrous Na2HPO4 6.42 g; NaH2PO4·H2O 2.03 g; KCl (5 mM final) 2.5 ml of 1 M stock; mannitol (50 mM final) 10 ml of 0.5 M stock; MgCl (15 mM final) 7.5 ml of 1 M stock
  3. Add good quality water to 500ml.
  4. Shake until everything has dissolved.
  5. Filter sterile into sterile falcon tubes and store in the fridge.