Lipo-UA Transfection Reagents

Cell transfection reagents facilitate the entry of nucleic acids into cells and enable the expression of the target gene within the cells. Liposomes are among the most commonly used and highly efficient transfection reagents in laboratories. Lipo-UA liposome cell transfection reagent is a highly effective DNA transfection reagent, offering advantages such as high transfection efficiency, broad applicability, ease of use, and cost-effectiveness, making it suitable for DNA transfection experiments in most cell types.

The liposome cell transfection kit consists of liposome reagent L and transfection enhancer E, with the following specifications. The number of 96-well plates that can be transfected is calculated based on a transfection of 0.1 μg DNA per well, and the ratio of transfection reagents DNA, L, and E is calculated as DNA (μg) : E (μL) : L (μL) = 1:2:3.

1.At96Confucius(or other cell culture plates of various sizes)Seed the cells to be transfected at an appropriate density, per well.100mulCell,Incubate overnight.Cell density during transfectiontransfection efficiency/Cytotoxicity has a significant impact, and different cell lines require optimization; typically, for example,HEK293orHeLaCell recommendationDuring transfectionThe cell density is at80% left and right

2.Calculate the required amount based on the area of the transfected cells.DNATotal amount, transfection enhancerEand liposome reagentsLTotal reagent volume. Recommended.DNADosage:6Orifice plate, per hole2.5mug DNA,24Orifice plate, per hole0.5mug DNA, 96Orifice plate, per hole0.1mug DNA. RecommendedDNA,EandLReagent ratio as perDNA (mug):E (muL):L (muL)For1:2:3Prepare the transfection reagent. If significant cytotoxicity is observed, reduce the amount accordingly.LReagent

3.Transfection reagent preparation: The following is for transfecting one plate.96Hole experiment board, Per hole0.1mug DNAConfigure as an example1 mLTransfection reagent. TransfectionOther cell culture plates of various sizes are according toDNAAdjust the amount of transfection reagent added to the culture medium. For example,96Hole experiment board100muLThe culture medium is supplemented with the following prepared components:10muLTransfection reagent(Per hole0.1mug DNA), 24Per well of the experimental plate500muLThe culture medium is supplemented with the following prepared components:FiftymuLTransfection reagent(Per hole0.5mug DNA),6Per well of the experimental plate2500muLThe culture medium is supplemented with the following prepared components:250muLTransfection reagent (Per hole2.5mug DNA)

a)use Opti MEM IDilute the nucleic acid to20mug/mL, take500μL,add20μLtransfection enhancerE, mix well and incubate at room temperature5 min

b)take500μL Opti MEM I, add30μLliposome reagentL, mix well

c)add all of the liposome reagentbprepared in stepLto the nucleic acid and transfection enhanceramixture prepared in stepE, mix well, and incubate at room temperaturefor 20 min

4.drop the transfection reagent prepared in step3into the corresponding cell culture wells, adding10μLper well, and gently shake a few times to mix

5.according to experimental needs, after transfection for a certain period of time, such as24 hr–96 hrdetect the expression of the target gene

It is not recommended to mix different batches.