Venor®GeMqOneStepisamycoplasmaqPCRdetectionkitwhichincludesallreagentsrequiredfortheqPCRreaction.Primer,nucleotides,polymeraseandtheinternalamplificationcontrolareprovidedready-to-useinalyophilizedreactionmix.RehydrationbufferandlyophilizedPositiveControlDNAarealsoprovidedinthekit.The assaycanbeperformedwithanytypeofreal-timePCRcyclerabletodetectthefluorescencedyesFAM™andHEX™.Theprotocolprovidedis recommendedforthefastandreliablescreeningofcellculturesupernatants.Thedetectionprocedurecanbeperformedwithin3hours.
MycoplasmasarespecificallydetectedbyamplifyingahighlyconservedrRNAoperon,ormoreprecisely,the16SrRNAcodingregioninthemycoplasmagenome.Themycoplasma-specificamplificationisdetectedat520nm(FAM™channel).The qOneStepPCRreactionmix includesprimersetsandFAM™labeledprobeswhichallowthespecificdetectionofallMollicutespeciessofardescribedascontaminantsofcellculturesandmediacomponents.EukaryoticDNAisnotamplifiedbythisprimer/probesystem.FalsenegativeresultsduetoPCRinhibitorsorimproperDNAextractionaredetectedbytheinternalamplificationcontrolwhich isalready includedinqOneStepreactionmix.Theamplificationoftheinternalamplificationcontrolisdetectedat610nm(HEX™channel).
TypeofPCR
TaqMan®based qPCRAssaywithFAM™andHEX™labeledprobes
ScopeofApplication
Applicableinresearchand developmentfordirecttestingofcellculturesandBIOLOGicals.Intendedforresearchuseonly.Notrecommendedforclinicaldiagnosticsortestingofhumansamples.
KitComponents
Primersets,nucleotides,internalamplificationcontrolandpolymeraseatoptimizedconcentrations,lyophilizedandpre-castinonetube
RehydrationBuffer
LyophilizedPositiveControlDNA
PCRgradewater
PackageSizes
qOneStepPCRreactionmixes arepreparedinaliquotsof25reactionseach.
- Cat.No.11-9102525tests
Cat.No.11-91100100tests
Cat.No.11-91250250tests
Resultevaluation
cycler-based,real-timePCR
RequiredConsumables
PCRreactiontubes
Requiredlabdevices
qPCRcycler
Pipettingequipment
Microcentrifuge
ShelfLifeandStorage
Venor®GeMqOneStepcomponents canbestored at+2to+8°Cforatleast12months.Afterrehydration,reagentsmustbestoredat≤18°C.
EP2.6.7compliance
No