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Zeta life/Sirius DNA Transfection Reagent P400025、 P400150/0.75ml/P400075

Overview

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
Component96-well24-well12-well6-well
Adherentcells1–4×1040.5–2×1050.25–1×106
Opti-MEM®Medium25μL×450μL×4100μL×4150μL×4
Sirius TransfectionReagent1,1.5,2,2.5μL2,3,4,5μL4,6,8,10ul6,9,12,15μL
Opti-MEM®Medium125μL250μL500ul700μL
DNA(0.5–5μg/μL)2.5μg5μg10ug14μg
DilutedDNATotal25μL50μL100ul150μL
DilutedSirius TransfectionReagent25μL50μL100ul150μL
Incubatefor5minutesatroomtemperature.
Component96-well24-well12-well6-well
DNA-lipidcomplexperwell10μL50μL100ul250μL
FinalDNAusedperwell100ng500ng1000ng2500ng
FinalSirius TransfectionReagentusedperwell0.2–0.5μL1.0–2.5μL2.0-5.0ul5.0–12.5μL
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.

Properties

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.

Background

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.

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