1. Reduce contamination risk with a separate area in your lab just for real-time PCR experiments—create an “RNA Zone” for sample prep extraction, external sample preparation, reaction mixture set-up, and running the real-time PCR instrument.
  2. Use Ambion RNaseZap® RNase Decontamination Solution to get rid of RNases and other contaminants prior to same prep procedures.
  3. Perform a bioinformatic evaluation of your target sequence before submitting the sequence into our online custom TaqMan® Assay design tool.
  4. Before resuspension of your lyophilized TaqMan® Assays, spin down the tube to ensure that you are resuspending everything in the tube.
  5. Make aliquots of your resuspended TaqMan® Assays to avoid repeated freeze/thaw cycles.
  6. Prevent cross contamination with filter tips or positive displacement pipettes.
  7. Cap the wells for No Template Control (NTC) prior to adding any sample cDNA/DNA or external standards during the total reaction preparation.
  8. Add DNA or RNA to your wells last to minimize cross contamination.
  9. Keep the bottom of your plate clean—use a support base for 96-well plates or a lint-free wipe/cloth for 384-well plates.
  10. Centrifuge your PCR plate before loading into the instrument to remove bubbles and ensure all reaction components are at the bottom of the well.
  11. When setting up and running replicate wells, make sure that your sample names, detector names, and detector tasks are the same.

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