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FAQ - PURPLE NIMT® Transfection Reagent


Questions

Answers 

Q1 Is it possible to separate transfected cells from untransfected cells by using a magnet?   A1 No. More than 95% of the cells take up PURPLE nanoparticles regardless of transfection efficiency. This means that some of the cells may not become transfected with siRNA. If you then try to sort with a magnetic column, all cells that have taken up the particles will become sorted.
Q2 Is PURPLE working on my cell line?   A2 View the following table to find all cells that we know have been tested with PURPLE.
Q3 Is PURPLE toxic to cells?   A3 If you optimize the transfection according to protocol, PURPLE is not toxic.
Q4 I would like to transfect cells ex vivo and then transplant cells into an animal. Is it possible to track cells with MRI?   A4 All NIMT products are based on superparamagnetic nano particles and can therefore be visualized with MRI.
Q5 Is it possible to dilute PURPLE in serum free medium, growth medium or buffers instead of water?   A5 No. To achieve optimal transfection efficiencies and to avoid toxic flocculations of the particles, you should always use double distilled water when you dillute the particles and siRNA prior to transfection.
Q6 Is it possible to store and reuse PURPLE particles?   A6 No. PURPLE should always be diluted just prior to use. 
Q7 When I add the transfection complexes to the cells, the medium becomes diluted with water. How does that affect the cells?   A7 The cell proliferation may become affected negatively to a small degree. If desired, the medium may be changed 6 hours after transfection, but the efficieny of transfetion may drop slightly.
Q8 Are there any cell specific transfection protocols?   A8 No. To get the best transfection efficiencies it is better to do an optimization using your own cells, plate format and type of siRNA. All instructions include an optimization protocol for adherent and suspension cell lines. We will gladly assist you to develop an optimization protocol, just click here to contact us.   
Q9 What is the lowest siRNA concentration that can be used with NIMT® PURPLE?   A9 The lowest concentration that can be used depends on the cell line. If it is a hard-to-transfect cell then a higher concentration of siRNA must be used (30-50 nM). An easy-to-transfect cell may only need 1-5 nM of siRNA.
Q10 How long should cells incubate before knockdown effects can be seen?   A10 Full knockdown can be measured as early as 8 hours after transfection.       
Q11 Is it possible to co-transfect DNA and siRNA with NIMT®transfection reagents?      A11  We would recommend to first transfect your cells with DNA using YELLOW and NIMT®Booster. Incubate the cells for approximately 6 hours, change medium and then transfect cells with siRNA using PURPLE.