Hi.As a part of my project I need to transfect SKBR3 and BT474 cell line, but these two are a little difficult to work with, especially when you don’t have lipofectamine! Does anybody here have the experience of working with another transfection reagent?
A few things; with SKBR3 it's best to use a pre-optimized transfection reagent (https://altogen.com/product/sk-br3-transfection-reagent-breast-adenocarcinoma/) that will help prevent any adverse effects from a generic protocol. With regards to cell confluency, it should never go above 80% before transfection, otherwise you won't be able to transfect all of the cells.
Hello Zahra,
You can use polyethyleneimine (PEI) for your transfections. It is very cheap and effective. You can try different concentrations of PEI.
Another option for transfection of mammalian cells is to use electroporation, but you are going to need an electroporator.
Good luck
hi
I agree with Ozen . critical point in DNA transfection has High quality Maxiprep (C:1ug/ul * OD 260/280 :1.8-2 ) and happy cells in log phase (confluency 80%) . Its better for stable DNA transfection use Linearized plasmid .
1- Phosphate calcium ( as harsh method )
2- Lipofectamin2000 ( liposomal base ) "FBS and antibiotic sensitive "
3- PEI ( cationic polymer base ) < such as Jetpei >
4- Electroporation ( Electrical shock base )
5- lentiviral particles ( Viral base )
good luck
http://www.abcam.com/index.html?pageconfig=resource&rid=11471
http://www.polyplus-transfection.com/products/jetpei/
https://www.theseus.fi/bitstream/handle/10024/48795/MERJALLE_2.pdf?sequence=1
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