Sirius DNA Transfection Reagentis a multi-component reagent that forms a complex with DNA and siRNA , then transports the complex insect cells.
Storage:Store at 4°C (Don’t Freezing).
Functional analysis:Cos 7 cells are transfected in 96 well plates with a reporter gene vector DNA, using 0.2-0.5 μlSirius Transfection Reagent and 0.1 μg DNA or 0.25 μl
Sirius Transfection Reagentand 0.1 μg DNA. Reporter gene activity is monitored via a colorimetric test against a reference lot. Activity must be 70% - 90%
compared to a reference lot in at least one of the tested amounts, listed above.
| ProcedureDetails | ||||
| Component | 96-well | 24-well | 12-well | 6-well |
| Adherentcells | 1–4×104 | 0.5–2×105 | 0.25–1×106 | |
| Opti-MEM®Medium | 25μL×4 | 50μL×4 | 100μL×4 | 150μL×4 |
| Sirius TransfectionReagent | 1,1.5,2,2.5μL | 2,3,4,5μL | 4,6,8,10ul | 6,9,12,15μL |
| Opti-MEM®Medium | 125μL | 250μL | 500ul | 700μL |
| DNA(0.5–5μg/μL) | 2.5μg | 5μg | 10ug | 14μg |
| DilutedDNATotal | 25μL | 50μL | 100ul | 150μL |
| DilutedSirius TransfectionReagent | 25μL | 50μL | 100ul | 150μL |
| Incubatefor5minutesatroomtemperature. | ||||
| Component | 96-well | 24-well | 12-well | 6-well |
| DNA-lipidcomplexperwell | 10μL | 50μL | 100ul | 250μL |
| FinalDNAusedperwell | 100ng | 500ng | 1000ng | 2500ng |
| FinalSirius TransfectionReagentusedperwell | 0.2–0.5μL | 1.0–2.5μL | 2.0-5.0ul | 5.0–12.5μL |
| Incubatecellsfor1–3daysat37°C.Thenanalyzetransfectedcells. | ||||
analysis is tested for viability.
Sirius DNA Transfection Reagentcomplexes must be made in serum-free medium such as Opti-MEM Reduced serum Medium and can be added directly to cells in
culture medium, in the presence or absence of serum .
It is not necessary to remove complexes or change and add medium after transfection.The amount of Sirius Transfection Reagentrequired for successful
transfection varies depending on the cell type and passage number. Start any new transfection by testing the recommended four concentrations of Sirius DNA to
determine an optimum amount.
Sirius DNA Transfection Reagent is a proprietary formulation for transfectin nucleic aicids into a wide range of eukaryotic cells . It is designed to transfect a
wide range of eukaryotic cells including insect cells and many cell lines not transfected well by other reagents （e.g.,MCF-7, RAW 264.7, PC-3, HeLa, MA-10,
HepG2, SH-SY5Y, A7r5, STO, SCC-61, STSAR-90, SQ20B, T98）.
Note: For the most up-to-date list of cell types that have been successfully transfected with Sirius DNA Transfection Reagent for life science research only.
