BioBharati LifeScience Pvt. Ltd.
Best Products for Quality Research of National Significance
EN-24, Kolkata Biotech Park, Salt Lake, Sector-V, Kolkata-700091
E-mail: biobharati@gmail.com, Web: www.biobharati.com
Cell No. 09836508080, Lab No. 033-40077640
PEiRfect Transfection Reagent:
Cat# BB-PT001
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
(see attached PDF), we are able to transiently transfect a number of cell-lines up to 40 – 90%.
Transfection of 293T cells with GFP using GG and AH PEI. Cells were treated with different DNA:PEI ratios and
different DNA/PEI incubation times at room temperature. 293T cells transfected with GFP using lipofectamine
was run as a control.
PEI Transfection Protocol
(Note – Protocol is written for a 2 cm plate. For larger or smaller transfections, scale up or down as necessary.)
Day 0
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Split cells to desired confluency.
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Add 50uL of serum-free, antibiotic-free DMEM to a 1.5mL eppitube
Add 2ug of DNA to serum-free, antibiotic-free DMEM
Vortex briefly
Add 4uL 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 50uL DNA-PEI mixture to the well drop wise
Swirl to distribute to mixture evenly throughout the well
Day 1
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Day 2
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After 6-8 hrs post-transfection, add fresh media (with serum and antibiotic) to the cells (this step is not
critical)
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Use or lyse cells for experiment
Day 3
Storage:
Stock solution: 1 mg/ml
 Make aliquot and store at -80°C; a working stock can be kept at 4°C. Don’t freeze the aliquot if once
thawed.