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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