PEiRfect Transfection Reagent

Cat# BB-PT001A (500µl) 
Cat# BB-PT001B (1000µl) 
Cat# BB-PT001C (2ml) 
Cat# BB-PT001D (5ml)

Description: Polyethylenimine is a cost-effective transfection reagent. In search for a cheap and effective transfection reagent BBL uses the positively charged polyplex forming compound polyethylenimine (PEI). This compound is commercially available from BBL as a non-modified chemical reagent. Using our optimized protocol, we are able to transiently transfect a number of cell-lines up to 40 – 90%.

Process:                      

(Note – Protocol is written for a 2 cm plate. For larger or smaller transfections, scale up or down as necessary.)

Day 0

• Split cells to desired confluency (70-80%).

Day 1

• Add 50µL of serum-free, antibiotic-free DMEM to a 1.5mL eppitube
• Add 2µg of DNA to serum-free, antibiotic-free DMEM
• Vortex briefly
• Add 4µL of PEI to the DNA-DMEM solution to obtain a final 1:2 DNA: PEI ratio (e.g. if you have 10ug of DNA, add 20uL of PEI, etc.)
• Vortex briefly
• Incubate at room temperature for 15 minutes
• Meanwhile, add fresh media to cells (with serum and antibiotic)
• After 15 minutes, add the 50µL DNA-PEI mixture to the well drop wise
• Swirl to distribute to mixture evenly throughout the well

Day 2

• After 6-8hrs post-transfection, add fresh media (with serum and antibiotic) to the cells

Day 3

• Use or lyse cells for experiment

Storage:

Stock solution: 1 mg/ml

Make aliquot and store at -20°C; a working stock can be kept at 4°C. Don’t freeze the aliquot if once thawed.  Exp. Date: 18 months upon receiving at proper storage condition as mentioned in datasheet.