FAQ: TargetAmp™ Kits

TargetAmp™: Selection Guide | Process | Performance | FAQ | Poster (3 MB PDF)

  1. The aRNA purification steps read "remove any remaining solution from the column using a vacuum apparatus". If we do not have a vacuum apparatus, could we use a longer centrifugation instead?
  2. How long does a TargetAmp reaction take?
  3. If I perform the in vitro transcription reactions using a thermocycler, can I add a 4°C overnight "hold" and continue with the aRNA purification the following day?
  4. What is the best way to purify my aRNA or aminoallyl-aRNA after I have completed the one amplification round or two amplification round procedures?
  5. At which steps in the TargetAmp RNA amplification process can I stop?
  6. What is the size of the aRNA (aminoallyl-aRNA) produced in a TargetAmp reaction?
  7. Why is it important to allow the AmpliScribe in vitro transcription reagents to warm up to room temperature before setting up the reaction?
  8. In the TargetAmp 2-Round amplification protocol the volume is adjusted to 3 µl using speed vacuum just prior to the 2nd-round 1st-strand cDNA synthesis step. Can I use a MicroCon concentrator instead?
  9. In the TargetAmp 2-Round amplification protocol the volume is adjusted to 3 µl using speed vacuum just prior to the 2nd-round 1st-strand cDNA synthesis step. What will happen if I reduce the volume to less than 3 µl?
  10. In the TargetAmp 2-Round amplification protocol the volume is adjusted to 3 µl using speed vacuum just prior to the 2nd-round 1st-strand cDNA synthesis step. What will happen if I accidentally dry the reaction?
  11. Is it possible to scale up or scale down the TargetAmp RNA amplification reaction?
  12. Why does Epicentre recommend doing the second strand cDNA synthesis reaction at 65°C? Other commercial kits and published procedures use 16°C.
  13. Will there be a benefit if I increase the in vitro transcription reaction times?
  14. What is the best RNA purification method to use to get total RNA for the TargetAmp Kit?
  15. Why do you recommend using SuperScript II and SuperScript III Reverse Transcriptases in the TargetAmp process?
  16. Can I use the TargetAmp Kits to amplify RNA isolated from laser-captured (or other microdissected) cells?
  17. When should I use a TargetAmp 1-Round kit or a TargetAmp 2-Round kit?
  18. What yield of aRNA (aminoallyl-aRNA) can I expect from a TargetAmp reaction?
  19. Do you have any citations that show the efficacy of RNA amplification from a single cell?
  20. I am currently performing two-rounds of RNA amplification. Can I get the same or better yield of aRNA (aminoallyl-aRNA) using a TargetAmp 1-Round Kit?
  21. Can I use the TargetAmp Kits for producing Cy-labeled aRNA?
  22. How much non-specific amplification product (background) will I get from a TargetAmp amplification reaction?
  23. Can the aRNA (aminoallyl-aRNA) produced by a TargetAmp reaction be used with either oligo arrays or cDNA arrays?
  24. What is the advantage of producing aminoallyl-aRNA rather than direct incorporation of a biotin-NTP, Cy-NTP or other fluorescent dye-NTP into the in vitro transcription reaction?