www.roche-applied-science.com

X-tremeGENEHPDNATransfectionReagent

Fortransientandstabletransfectionofeukaryoticcells

Version05

Contentversion:May2011

Cat.No.06365752001

Cat.No.06366244001

Cat.No.06366236001

Cat.No.06366546001

Trial-pack

0.4ml

1ml

5×1ml

Storeat–15to–25°C

1.

WhatthisProductDoes

NumberofTests

Usingthestandardprocedure,1mlofX-tremeGENEHPDNATrans-

fectionReagentcanbeusedtoperformupto10,000transfectionsin

96-wellplates.

Formulation

X-tremeGENEHPDNATransfectionReagentisaproprietaryblendof

lipidsandothercomponentssuppliedin80%ethanol,filteredthrough

0.2

␮

mporesizemembrane,andpackagedinglassvials.Itdoesnot

containanyingredientsofhumanoranimalorigin.

StorageandStability

StoreX-tremeGENEHPDNATransfectionReagentat–15to–25°C,

withthelidtightlyclosed.Thereagentisstableuntiltheexpiration

dateprintedonthelabelwhenstoredundertheseconditions.

L

X-tremeGENEHPDNATransfectionReagentremainsfullyfunc-

tionalevenafterrepeatedopeningofthevial(atleastfivetimes

overatwo-monthperiod),aslongasthevialistightlyrecapped

andstoredat-15to-25°C.

L

Notethattheshippingtemperatureofthisproductisdifferent

fromthestoragetemperature.Thesedifferenttemperatureswill

notaffectproductperformanceorproductstability.

SpecialHandling

N

Afterremovingtheamountrequired,tightlyclosethevialwiththe

lidimmediatelyafteruse.

N

Alwaysbringthevialto+15to+25°CandmixX-tremeGENEHP

DNATransfectionReagentpriortoremovingtheamountrequired

vortexingforonesecond.

N

DonotaliquotX-tremeGENEHPDNATransfectionReagent;store

intheoriginalglassvials.

N

MinimizethecontactofundilutedX-tremeGENEHPDNATrans-

fectionReagentwithplasticsurfaces.

N

Foruse,theminimumamountofX-tremeGENEHPDNATransfec-

tionReagent:DNAcomplexis100µl.Complexformationatlower

volumescansignificantlydecreasetransfectionefficiency.

N

Donotusetubesormicroplatesmadeofpolystyrenefor

X-tremeGENEHPTransfectionReagent:DNAcomplexprepara-

tion.Whennotabletoavoidpolystyrenematerials,makecertainto

pipetthetransfectionreagentdirectlyintotheserum-freemedium

(

e.g.,

Opti-Mem).

N

Donotusesiliconizedpipettetipsortubes.

AdditionalEquipmentandReagentsRequired

Additionalreagentsandequipmentrequiredtoperformtransfection

assaysusingX-tremeGENEHPDNATransfectionReagentinclude:

➤

StandardLaboratoryEquipment

.

Ϫ

Standardcellcultureequipment(

e.g.,

biohazardhoods,incuba-

tors)

Ϫ

Standardpipettesandmicropipettes

Ϫ

Vortexmixer

➤

ForPlasmidPreparation

Ϫ

Purifiedplasmidstock(0.1–2.0µg/µl)insterileTE(10mMTris,

1mMEDTA,pH8.0)bufferorsterilewater

Ϫ

GenopurePlasmidMidiKit*orGenopurePlasmidMaxiKit*to

prepareplasmid

➤

ForVerificationofVectorFunction

Ϫ

Assayappropriatelyfortransfectedgene

Ϫ

G-418Solution*orHygromycinB*(optionalforstabletransfec-

tionexperiments)

➤

ForTransfection-ComplexFormation

Ϫ

Opti-MEMIReducedSerumMediumorserum-freemedium

Ϫ

Sterilepolypropylenetubesorround-bottom96-wellplates

➤

GrowingCells

Ϫ

Selectsubconfluentculturesinlogphaseforpreparationofcell

cultures

Ϫ

Quantifycellnumbertoreproduciblyplatethesamenumberof

cells

Application

X-tremeGENEHPDNATransfectionReagentisahighperformance

transfectionreagent,freeofanimal-derivedcomponents.Benefitsof

X-tremeGENEHPDNATransfectionReagentinclude:

•

Designedtotransfectabroadrangeofeukaryoticcells,including

insectcells,manycelllinesnottransfectedwellbyotherreagents,

andhard-to-transfectcelllines(

e.g.,

HT-1080,K-562,HepG2).

•

Canbesuccessfullyusedinavarietyofapplications,suchasgene

expressionanalysisandproteinproductionusingtransientlytrans-

fectedcells,generationofstablecelllines,expressionofshRNAfor

geneknockdownstudies,drugdiscoveryprograms,andtarget

evaluation.Samplesanddetailedtransfectionprotocolsareavail-

ableat

http://www.powerful-transfection.com.

•

Producesminimalcytotoxicityorchangesinmorphologywhenade-

quatenumbersofcellsaretransfected,eliminatingtherequirement

tochangemediaafteraddingthetransfectioncomplex.

•

Suitablefortransientandstabletransfection.

•

Functionsverywellinthepresenceorabsenceofserum.

Forlifescienceresearchonly.



Notforuseindiagnosticprocedures.

0511.

06479774001

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