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Transcription-Mediated Amplification (TMA)
Transcription-Mediated Amplification (TMA) uses two enzymes, Reverse Transcriptase,
and RNA polymerase, to produce billions of copies of RNA amplified target from the
purified target nucleic acid.
The amplification process takes approximately one hour.
1. The MTU is transferred to the AMP load station where Amplification Reagent is
pipetted into each tube and mixed.
2. The MTU is transferred back to the High Temperature Incubator for the primer anneal
step, which promotes the binding of the primers to the target nucleic acid.
3. The MTU moves to the Transition Incubator to cool before Enzyme Reagent addition.
4. The MTU moves to the AMP load station. The pipettor adds Enzyme Reagent. The
MTU is mixed and transferred to the AMP/RT Incubator for amplification at 42.7 °C.
a. Reverse Transcriptase uses the primers to create a DNA copy of the target RNA.
If the target is DNA, the process skips to Step d.
b. The RNase H activities of reverse transcriptase degrade the original target RNA.
c. The second primer binds to the new DNA copy and reverse transcriptase creates
a double-stranded DNA duplex.
d. The RNA polymerase, through transcription, makes multiple RNA copies of the
DNA template.
e. The process repeats until the reagent supply is exhausted.
5. The amplified RNA, called "amplicon," is ready to be hybridized with the labeled
nucleic acid probe in the Hybridization Protection Assay.
Hybridization Protection Assay (HPA)
The Hybridization Protection Assay (HPA) process hybridizes the amplicon to single-
stranded nucleic acid probes that are labeled with an acridinium ester (AE) molecule and
then it selects and detects the hybridized probes.
®
Panther System
Operator's Manual
AW-17791-001 Rev. 002 (EN)
Overview
27
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