Gene Expression Analysis |
RNA Amplification for qPCR
Order
| Protocol
| Citations
| Related Products
Applications
- Generation of large amounts of cDNA from small amounts of compromised
or degraded total RNA preparations for RT-PCR or qRT-PCR studies.
- Generation of large amounts of cDNA from small amounts of compromised
or degraded total RNA preparations for archival purposes.
|
 |
Figure 1 (click to enlarge). Overview of the MessageBOOSTER™ Whole-Transcriptome
Kit procedure. The kit amplifies the RNA from small amounts of compromised
or degraded total RNA and then converts the amplified RNA to cDNA that is
ready for qPCR. The single-tube reaction can be completed in 1 day. |
The MessageBOOSTER™ Whole-Transcriptome cDNA Synthesis Kit for
qPCR uses the proprietary MessageBOOSTER™ Whole-Transcriptome Primers
to prime cDNA synthesis. These primers anneal to and prime cDNA synthesis from
all types of RNA found in the cell (Fig. 1). Thus, this kit is ideal for amplifying
RNA and subsequently producing cDNA from compromised or degraded RNA samples
such as those obtained by laser-capture microdissection, cell-sorting methods,
or from formalin-fixed paraffin-embedded (FFPE) tissue.
Note: The ability to amplify RNA from FFPE samples using the MessageBOOSTER™ Whole-Transcriptome
cDNA Synthesis Kit is dependent on factors that are often difficult for the
user to control.
We have observed that some RNA samples isolated from FFPE tissue have been
successfully amplified and converted to cDNA while other samples have failed.
Therefore, your results may vary depending on the condition of the FFPE sample.
This kit will not amplify RNA from a single cell. Use the original MessageBOOSTER™ cDNA
Synthesis Kit for qPCR for single-cell amplification.
Benefits
- Significantly increase the number of qRT-PCRs that can be obtained from
small amounts of degraded RNA samples (Table 1).
- Produce cDNA with greatly improved 3´/5´ ratio from degraded
RNA samples.
- Use as little as 1 ng of degraded total RNA per reaction.
- One-day, single-tube reaction.
| Partially degraded |
10 ng |
>10 |
>500 |
| Severely degraded |
100 ng |
>10 |
>500 |
|
Table 1. The number of qPCR amplifications from cDNA produced by the
MessageBOOSTER™ Whole-Transcriptome Kit. |
*MessageBOOSTER Products are covered by intellectual property licensed
to EPICENTRE Technologies Corporation from Johnson & Johnson Pharmaceutical
Research & Development, L.L.C., and by intellectual property sublicensed
to EPICENTRE Technologies Corporation from Incyte Corporation.
MessageBOOSTER™ cDNA Synthesis Kit for qPCR
MessageBOOSTER™ cDNA Synthesis from Cell Lysates Kit
[top of page]
| 
