Transfection

Set up a reaction

Pick a format. DNA stays at the same micrograms per microliter when you change format, and PEI stays at 4× the DNA.

DNA per well or flask (I am asking about per well or per flask- Say it in µg/well) How many wells do you need to transfect ? Make sure- For example- Only 1 well or N well DNA stock (ng/µL)- It's in nanogram per ul. PEI stock (ug/uL). We have 1 ug per ul. So, for 1 ug DNA, use 4 ul of PEI.

Add 10% extra for a master mix

What to pipette

One 24-well well.

Volumes by format

Example DNA keeps the 24-well concentration: 1 µg in 130 µL. PEI is 4× that DNA. Pipette volumes assume a 1000 ng/µL DNA stock and a 1 mg/mL PEI stock.

Format	Area	Reaction	DNA	PEI	Pipette DNA	Pipette PEI
24-well	1.9 cm²	130 µL	1.00 µg	4.00 µg	1.0 µL	4.0 µL
6-well	9.6 cm²	260 µL	2.00 µg	8.00 µg	2.0 µL	8.0 µL
T25	25 cm²	677 µL	5.21 µg	20.84 µg	5.2 µL	20.8 µL
T75	75 cm²	2.03 mL	15.62 µg	62.48 µg	15.6 µL	62.5 µL
T150	150 cm²	4.06 mL	31.25 µg	125.00 µg	31.3 µL	125.0 µL

Match these numbers to your own protocol before you add the complex to cells. Incubate the finished mix for whatever time that protocol uses, often 10–20 minutes at room temperature.

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