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C
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There is evidence to suggest that mycoplasma can survive in the absence of cells, some may even proliferate, but as a rule they are much more viable if cells are present.
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For optimal assay conditions, reconstituted reagent and substrate should be used fresh within 2 hours after reconstitution.During this time they can be kept at room temperature.For the MycoAlert® Kit: Unused, reconstituted components can be aliquoted...
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The standard protocol calls for taking 2 ml of culture media, spinning down any cells and removing 100 µl of supernatant as sample.Smaller initial aliquots of media may be taken to start with if necessary.
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N
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™
?
Specific blood cell populations can be enriched/purified for Nucleofection™ by MACS™ selection Kits (Miltenyi Biotec GmbH), RosetteSep™ Kits (StemCell Technologies Inc.) or Dynal™ beads (Invitrogen Corporation).
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N
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We recommend using negative selection (depletion) because your purified cells are "untouched", not influenced, and strictly not activated.Positive selection may cause increasing amounts of dead cells and could also lead to activation of the cells due...
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?
Yes. To check the quality of your DNA we strongly recommend determining the A260:A280 ratio. It should be ideally 1.8 - 2.0 for a good DNA preparation, but least 1.6.
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?
No, the MycoAlert® Mycoplasma Detection Assay (as well as the MycoAlert® PLUS Mycoplasma Detection Assay) is not quantitative; it has been designed to give a simple Yes or No answer, pertaining to the presence of Mycoplasma.
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The molecular weight is approximately 2.3x10e6 g/mol based on the average MW of a base pair being 660 g/mol.
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It is highly recommended that the samples are spun down prior to testing. If cells are present in the assay they will contribute to the initial reading producing a higher background. This in turn can decrease the difference between the background and...
C
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Yes. You should verify the integrity of your DNA on an agarose gel to see if it is degraded. Compare undigested plasmid to plasmid DNA digested with a suitable single cut restriction enzyme to linearize. Supercoiled plasmid will run faster than...
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