jetPEI® DNA transfection, HTS application

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Do not add antibiotics to the Transfection Medium

Protocol tips
Do not add antibiotics to the Transfection Medium

Publication protocol

C. Cell transfection and clonal selection
MDA-MB-231, MDA-MB-435, and GILM2 transient and stable transfections were carried out using jetPEI reagent (PolyPlus Transfection, France) according to the manufacturer's instructions. In order to produce stable clones, a 1∶5–1∶20 dilutions of 5×106 transfected cells was performed into 100 mm Petri dishes 48 hours post transfection. Selection was commenced the day after. Selective media consisted of DMEM without phenol-red, supplemented with 5% dextran charcoal fetal bovine serum (FBS), 4 mM L-glutamine, antibiotics (10 units/ml of penicillin and 50 µg/ml streptomycin) and the selective drug. Selection of stable clones was performed at 0.4 mg/ml G418 (Calbiochem) for pCDNA3-neo based clones, or at 0.2 mg/ml Hygromycin B (A.G. Scientific) for pIREShyg3 and pCMV-Bam-ERα-HygroR based clones. Selective media was refreshed every 3 days thereafter. When colonies were big enough and interspaced, they were transferred to 48- well cell culture plates. For long term maintenance, 0.2 mg/ml G418, or 0.1 mg/ml Hygromycin B were used.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA transfection Mammalian cells - Immortalized cell lines MDA-MB-231 using jetPEI® DNA transfection, HTS application from Polyplus transfections.

Manufacturer protocol

Download the product protocol from Polyplus transfections for jetPEI® DNA transfection, HTS application below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA transfection Mammalian cells - Immortalized cell lines MDA-MB-231 using jetPEI® DNA transfection, HTS application from Polyplus transfections. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.