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.

Cytotoxicity analysis:Cell growth is assessed using theCell Counting Kit (K009,ZETA LIFE Inc.) whichis an indirect measure of cytotoxicity. The remainder of cells from functional

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.