Transfection Reagent FAQs
1. What kind of reagent is MegaTran 2.0?
MegaTran 2.0 is a biodegradable polymer based DNA transfection reagent specially designed for the effective and reproducible transfection on HEK293, COS-7, NIH-3T3, HeLa, CHO and a broad ranges of hard-to-transfect mammalian cells. A remarkable feature of the reagent is the rapid and complete degradation of polymer after transfection complex endocytosis, leading to much less cytotoxicity.
2. What cell lines have been tested for use with MegaTran 2.0?
Based on our innovative polymer synthesis technology, MegaTran 2.0 is formulated to be a powerful transfection Reagent that ensures effective and reproducible transfection with less cytotoxicity. MegaTran 2.0 was shown to deliver genes to various established cell lines as well as primary cells, including CHO, HEK293, HepG2, HeLa, COS-7, NIH3T3 et al. Please contact our Technical Support Team regarding your cells.
3. Should I use the same DNA:MegaTran ratio for cell types that have not been tested?
For different cell types, the optimal ratio of MegaTran 2.0 (µL):DNA (µg) is around 3:1. We recommend the MegaTran 2.0 (µL):DNA (µg) ratio of 3:1 as a starting point which usually gives satisfactory transfection efficiency with invisible cytotoxicity. To ensure the optimal size of MegaTran 2.0/DNA complex particles, we recommend using serum-free DMEM with High Glucose to dilute DNA and MegaTran 2.0.
4. What level of efficiency should I expect from MegaTran 2.0?
When HEK293 and CHO cells are used, over 80% of the cells can be transfected. However, with different cell lines and different DNA preparation, the efficiency varies. Each of them needs to be optimized individually.
5. What level of toxicity should I expect?
MegaTran 2.0 display much less toxicity compared to the commonly used Lipofectamine. With the recommended protocol using CHO cells, the transfected cells appears to be healthy morphologically and continue to produce the transgene protein even at day 6 post transfection.
6. Is the transfection efficiency sensitive to cell density?
Yes. The best transfection efficiency is usually achieved with 80% confluent cells. Different cell lines need individualized optimization for the best condition.
7. Will antibiotics in the growth medium interfere with MegaTran 2.0 transfection?
High serum levels (>5%) with antibiotics usually do not have inhibitory effect on transfection efficiency. We recommend using complete serum/antibiotics-containing medium as a starting point. For maximal efficiency and lower cytotoxicity, perform transfection on cells with high density. We recommend transfecting on cells with ~80% confluency.
8. Do I need to replace the media after MegaTran 2.0 transfection?
Remove MegaTran/DNA complex-containing medium and replace with fresh complete serum/antibiotics containing medium 12~18 hours post transfection. For sensitive cells, to lower cytotoxicity, remove MegaTran/DNA complex and replace with complete medium 5 hours after transfection.
9. What are the storage conditions and stability of MegaTran 2.0 transfection reagent?
We recommend the storage condition as 40C. It is stable for up to 12 months from date of receipt under proper storage and handling conditions.
10. What is the recommended method for the quantitation of my DNA?
We recommend using UV spectrophotometers to obtain the accurate quantity and the purity of the DNA samples.
11. Where can I read more about this reagent?
Please access the product documentation and data on our website at the following address:.
12. How should I cite your product?
Here is our guideline:
Full Product Name, OriGene Technologies Inc., Rockville, MD, USA, Catalog #, Lot #.
In addition, we would love to hear from you when the paper is published. Email us a copy of the accepted manuscript and receive a special gift.