产品介绍

       The  Mycoplasma test kit   offers  high  specificity  and  sensitivity  to  minimize  false positives  while  ensuring  coverage  over  2 0 0   strains  of  Mycoplasmas  for  quick  and  reliable routine screening of cell cultures less than  1  hour. Mycoplasmas are highly undesirable,  easily acquired  and  notorious  for an  elusive  onset  of infection.  It can  alter the  infected  cells at  a molecular  level  leading  to  visible  changes  in  cell  morphology  and  growth  characteristics.  Timely  detection  of  Mycoplasmas  in  cell  cultures  is  recommended  in  order  to  deter  wide-  spread  contamination  and  save  on the  costly  efforts  of  elimination.  The  MasterMix  contains gel loading dye,  making it convenient to  load for gel electrophoresis.

Protocol:
1.Cell should remain in culture for at 48-72 hours undisturbed prior to screening and is at least 80% confluence.     Collect 2.5 ul directly from culture.
2.Mix individual components  before  use,  and assemble reaction on ice.
       

3.Gently mix the reaction and briefly centrifuge. Run thermocycling conditions for standard PCR:

4.After PCR,  maintain the reaction at 4° C or store at -20 ° C until use.

5.Analyze  the  amplification  products  by  agarose  gel  electrophoresis.

6 .Visualize by ethidium bromide or SafeView   ( Cat  No.  K108) staining.

General 

•   PCR product approximately 500 bp in length indicates that the cell culture tested is contaminated with Mycoplasma. Note that the length of the PCR product will vary between 370-550 bp depending on the different Mycoplasma strains.

•   Keep reaction mixture on ice prior to running the PCR, and start the PCR as soon as the reaction mixture is prepared.

Sensitive Detection

KEL's Mycoplasma PCR Detection Kit can detect samples containing as low as 10 genome copies of Mycoplasma. KEL's Mycoplasma PCR Detection Kit (Cat. No. KC331-01) was used to detect samples containing ~10 to 100,000 genome copies of Mycoplasma. Presence of Mycoplasma was confirmed as a 450 bp band.