3rd July 2024

MessageBOOSTER™ cDNA Synthesis Kit for qPCR

Applications

  • Significantly increasing the number of qRT-PCRs that can be obtained from very small amounts of total RNA, as little as one cell (10 pg).
  • Generation of large amounts of cDNA from very small samples of intact total RNA for archival purposes.

The MessageBOOSTER™ cDNA Synthesis Kit for qPCR* enables the user to perform sensitive qPCR amplifications using the total RNA from very small populations of cells, even from as little as one cell (Table 1).

The kit amplifies the mRNA contained in a small total RNA sample and then converts the amplified RNA to cDNA that is ready for PCR or qPCR (Fig. 1). The MessageBOOSTER cDNA Synthesis Kit for qPCR uses oligo(dT) to prime cDNA synthesis. Therefore, this kit is best used with intact total RNA preparations.

Benefits

  • Even low-abundance transcripts from total RNA of one cell are readily detected (CT values <35 cycles) (Fig. 2).
  • Collect small RNA samples less often.
  • Reactions can be performed directly from whole-cell lysates without the need for purifying RNA.1
  • Perform a MessageBOOSTER cDNA Synthesis Kit reaction in 1 day.
  • The linear RNA amplification and cDNA synthesis processes preserve the gene expression profile.

Figure 1 (click to enlarge). Overview of the MessageBOOSTER™ cDNA Synthesis Kit procedure. The kit uses a linear RNA amplification process to amplify the RNA from a small population of cells, then converts the amplified RNA to cDNA that is ready for PCR. The procedure can be completed in 1 day.

 

Figure 2. Sensitivity of the MessageBOOSTER™ cDNA Synthesis Kit. cDNA produced from 10 pg of total NRK RNA significantly improved the sensitivity (lowered the CT) of detecting the low-abundance PBGD transcript compared to cDNA produced from 10 pg of unamplified RNA.

 

Amount of Total RNA in a MessageBOOSTER™ Reaction Number of qPCRs That Can Be Performed
Low- to Medium- Abundance Transcriptsa Medium- to High- Abundance Transcriptsa
10 pg (~1 cell) ≥10 ≥100
100 pg (~10 cells) ≥100 ≥1,000
500 pg (~50 cells) ≥500-1,000 ≥5,000-10,000
Table 1. The number of qPCR amplifications that can be performed using cDNA produced by a MessageBOOSTER™ cDNA Synthesis Kit. The number of qPCRs is dependent on the amount of input total RNA and the abundance of the transcript(s) of interest.
Low-Abundance Transcripts = 1-1,000 copies per cell
Medium-Abundance Transcripts = 1,000-10,000 copies per cell
High-Abundance Transcripts = 10,000-100,000 copies per cell

 

Product Citations

  1. Grunenwald, H. et al. (2006) Epicentre Forum 13(3), 22.
  2. DeFazio, R.A. et al. (2006) J. Neuroscience 26, 3971.
  3. Kaeffer, B. et al. (2007) Pediatric Research 62, 564.
  4. Lochner, M., et al. (2008) J. Exp. Med. 205, 1381.
  5. Graham, D.M. et al. (2008) J. Neurophysiology, 99, 2522.
  6. Liu, X. et. al. (2009) Am. J. Physiol. Lung Cell Mol Physiol. 296, L158
  7. Yakovlev, I.A. et al. (2008) Fungal Genetic and Biology 45, 498
  8. Jung, Y. et al. (2008) Stem Cells Express 10, 1634
  9. Xi, D. et al. (2008) J. Neurosci Method 10, 1016
  10. Michel, M-L. et al. (2008) Proc. Nat’l Acad Sci-USA 105(50), 19845
  11. Satoh-Takayama, N. et al. (2008) Immunity 29, 958
  12. Bouskra, D. et al. (2008) Nature 10, 1038
  13. Yakovlev, I.A. et al. (2008) Fungal Genetics and Biology 45, 498
  14. Roberts, C.D. et. al. (2009) J. Physiology 587, 1657
  15. Peduto, L. et. al. (2009) J. Immunology 182, 5789
  16. Xi, D. et. al. (2009) Int’l J. Neuropsychopharmacology, May 13:1-14

*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.

Ordering

Catalog No. Size
MessageBOOSTER™ cDNA Synthesis Kit for qPCR
MB060124 24 Reactions
Contents: MessageBOOSTER™ T7-Oligo(dT) Primer A, RNase Inhibitor, MessageBOOSTER™ Reverse Transcription PreMix, MessageBOOSTER™ DNA Polymerase PreMix 1, MessageBOOSTER™ DNA Polymerase 1, MessageBOOSTER™ cDNA Finishing Solution, MessageBOOSTER™ Random Primers, MessageBOOSTER™ RNase H, MMLV Reverse Transcriptase, MessageBOOSTER™ In Vitro Transcription PreMix A, MessageBOOSTER™ T7 RNA Polymerase, MessageBOOSTER™ T7 Transcription Buffer, RNase-Free DNase I, Forward and Reverse Control PCR Primers, DTT, RNase-Free Water, Poly(I), NRK Total RNA Control.