Venor®GeMqEPutilizesquantitative,real-timePCR(qPCR).Thekitcanbeperformedwithanytypeofreal-timePCRcyclerabletodetectthefluorescencedyesFAM™andHEX™.Theprotocolprovidedispreferredforfastandreliablescreeningofcellculturesupernatantsmostapplicableinresearchanddevelopment.Thedetectionprocedurecanbeperformedwithin3hours.
MycoplasmasarespecificallydetectedbyamplifyingahighlyconservedrRNAoperon,ormorespecifically,the16SrRNAcodingregioninthemycoplasmagenome.Themycoplasma-specificamplificationisdetectedat520nm(FAM™channel).ThekitincludesprimerandFAM™labeledprobeswhichallowthespecificdetectionofallMollicutesspeciessofardescribedascontaminantsofcellculturesandmediacomponents.EukaryoticDNAisnotamplifiedbythisprimer/probesystem.
FalsenegativeresultsduetoPCRinhibitorsorimproperDNAextractionaredetectedbytheinternalamplificationcontrolwhichcanbeaddedtothePCRmastermix.Theamplificationoftheinternalamplificationcontrolisdetectedat610nm(HEX™channel).
TaqMan®based qPCRAssaywithFAM™andHEX™labeledprobes
Venor®GeMqEPisusedfordirectdetectionofMollicutes(Mycoplasma,Acholeplasma,Spiroplasma)contaminationincellculturesandcellmediacomponents.
lyophilizedprimer/nucleotide/probe/polymerase/mixinaliquotsof25tests
rehydrationbuffer
lyophilizedinternalamplificationcontrol
lyophilizedpositivecontrol
PCRgradewater
cyclerbased,real-timePCR
PCRreactiontubes
OptionalforprocessvalidationandEP2.6.7complianttesting:
InternalControlDNAextra(4vialsfor300µleachofinternalamplificationcontrol;Cat.No.11-9905)
10CFU™SensitivityStandardsavailableforallEP 2.6.7listedmycoplasmaspecies
qPCRcyclerwithFAM™andHEX™filter
variablemicroliterPipettes
benchtopcentrifugefor1.5mlreactiontubes
Componentscanbestoredat+2to+8°Cforatleast12months.Afterrehydrationthereagentsmustbestoredat-18°C.
Yes.
Pleasenotethatvalidationdataareprovidedforinformationpurposeonly.EP2.6.7clearlystates“Wherecommercialkitsareused…,documentedvalidationpointsalreadycoveredbythekitmanufacturercanreplacevalidationbytheuser.Nevertheless,theperformanceofthekitwithrespecttoitsintendedusehastobedemonstratedbytheuser(e.g.detectionlimit,robustness,cross-detectionofotherclassesofbacteria.”.Pleasefeelfreetocontactusifyouneedfurtherassistance.
Fig.AmplifieddilutionseriesofMycoplasmafermentans,performedona
CorbettRotorGene®6000.
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