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POLOdeliverer™3000TransfectionReagent
PRODUCTDESCRIPTION
POLOdeliverer™3000TransfectionReagentisasuperiorpolymer-basedformulationforthetransfectionofDNAintoeukaryoticcellsandprovidesthefollowingadvantages:
l Highesttransfectionefficiencyinmanycelltypesandformats
l Simpleprocedurewithoutmediumchange
l TransientandstabletransfectionofadherentandsUSPensioncells
l Verylowcytotoxicity
POLOdeliverer™3000givessuperiortransfectionefficiencyforabroadrangeofcelllines,includingandnotlimitedtothefollowingcelllines:
293 CHO-S(adherent) Hela HT29
293A COS-1(w/oserum) PC12 NIH3T3
293T COS7-L MDCK MCF7
PrimaryHumanFibroblasts(w/oserum) Vero HepG2
SHIPPINGCONDITIONS
POLOdeliverer™3000TransfectionReagentisshippedatroomtemperature.
STORAGECONDITIONS
POLOdeliverer™3000TransfectionReagentshouldbestoredat4ºC.
STABILITY
ThestabilityofPOLOdeliverer™3000rransfectionreagentisguaranteedfor6monthswhenithasbeenstoredasrecommended.POLOdeliverer™3000Reagentshouldnotbefrozen.
GeneralprotocolofplasmidDNAtransfectionwithPOLOdeliverer™3000
1. Onedaybeforetransfection,plate3×105cellsin2mlmediumperwellofa6-wellcultureplate.Incubateat37ºC(5%CO2)overnight.Theconfluencyofcellsshouldbe60-80%whentransfection.
2. Dilute2μgDNAin100μlmedium,suchasDMEM,RPMI1640,opti-MEM.
3. Dilute4μlPOLOdeliverer™3000in100μlthesamemedium.
4. AddPOLOdeliverer™3000solutiontotheDNAsolutiondropwiseandmixbyPipetteorvortex,thenincubatedatroomtemperaturefor15minutes(37ºCincubationfor5minutesofthePOLOdeliverer™3000solutionandtheDNAsolutionseparatelybeforemixingisrecommended).
5. Withoutchangingtheculturemedium,addthemixturedropbydropintothecultureandmixbygentlyrockingtheplate.
6. Maintainthetransfectedcellsat37ºC,5%CO2for24-48hourswithoutmediumchange(Ifyouwant,themediumcanbechanged12–24hoursaftertransfection.).
TableRecommendedconditionsfortransfectionwithPOLOdeliverer™3000*
Culturevessel | 96-well | 24-well | 12-well | 6-well | 35-mm | 60-mm | 100-mm | 150-mm | T25 | T75 |
Surfacearea(cm2) | 0.3 | 2 | 4 | 10 | 10 | 20 | 60 | 140 | 25 | 75 |
RecommendedvolumeofPOLOdeliverer™3000(μl) | 0.4 | 2 | 3 | 4 | 4 | 10 | 20 | 50 | 12 | 40 |
RecommendedqualityofDNA(μg) | 0.2 | 1 | 1.5 | 2 | 2 | 5 | 10 | 25 | 6 | 20 |
*Fortransfectionindifferentcelllines,optimizationofPOLOdeliverer™3000amount,DNAconcentration,cellnumberandtransfectiontimeisrecommended.
Productnameandcatalognumbers
Name | Size | Catalognumber |
POLOdeliverer™3000Reagent | 0.75ml | P30001 |
POLOdeliverer™3000Reagent | 1.5ml | P30002 |
POLOdeliverer™3000Reagent | 1.5ml*10 | P30003 |
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