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).
TaqMan®based qPCRAssaywithFAM™andHEX™labeledprobes
Applicableinresearchand developmentfordirecttestingofcellculturesandBIOLOGicals.Intendedforresearchuseonly.Notrecommendedforclinicaldiagnosticsortestingofhumansamples.
Primersets,nucleotides,internalamplificationcontrolandpolymeraseatoptimizedconcentrations,lyophilizedandpre-castinonetube
RehydrationBuffer
LyophilizedPositiveControlDNA
PCRgradewater
qOneStepPCRreactionmixes arepreparedinaliquotsof25reactionseach.
cycler-based,real-timePCR
PCRreactiontubes
qPCRcycler
Pipettingequipment
Microcentrifuge
Venor®GeMqOneStepcomponents canbestored at+2to+8°Cforatleast12months.Afterrehydration,reagentsmustbestoredat≤18°C.
No
SouthernBiotech/Goat Anti-Human Lambda-Alexa Fluor® 555/2070-32/1.0 mg
vectorlabs/Biotinylated Aleuria Aurantia Lectin (AAL)/B-1395/1 mg
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Mouse IgG)/PK-6102/1 kit
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Universal)/PK-6200/1 kit
vectorlabs/Unconjugated Musa Paradisiaca (Banana) Lectin (BanLec)/L-1410/5 mg
GiottoBiotech/Calmodulin N60D/5 mg/G02CLM60cn