Product FAQ

What can I do to improve the sensitivity of my qPCR assay?


If you are targeting a low-abundance gene, you may have trouble getting Ct values in a good, reliable range (Ct > 32). To increase the sensitivity of the assay, you may want to consider the following:

- Increase the amount of RNA input into your reverse transcription reaction, if possible
- Increase the amount of cDNA in your qPCR reaction (20% by volume max)
- Try a different reverse transcription kit, such as our SuperScript VILO Master Mix, for the highest cDNA yield possible
- Consider trying a one-step or Cells-to-CT type workflow (depending on your sample type)

Answer Id: E7510

Was this answer helpful?

Thank you for your response

Citations & References

Clinical and biological significance of CDK4 amplification in well-differentiated and dedifferentiated liposarcomas.

  • Authors: Italiano A, Bianchini L, Gjernes E, Keslair F, Ranchere-Vince D, Dumollard JM, Haudebourg J, Leroux A, Mainguene C, Terrier P, Chibon F, Coindre JM, Pedeutour F
  • Journal: Clin Cancer Res 2009 18:5696-5703
  • PubMed ID: 19737942

Citations & References

The effects of 4-methylumbelliferone on hyaluronan synthesis, MMP2 activity, proliferation, and motility of human aortic smooth muscle cells.

  • Authors: Vigetti D, Rizzi M, Viola M, Karousou E, Genasetti A, Clerici M, Bartolini B, Hascall VC, De Luca G, Passi A
  • Journal: Glycobiology 2009 5:537-546
  • PubMed ID: 19240269

Citations & References

Arabidopsis primary microRNA processing proteins HYL1 and DCL1 define a nuclear body distinct from the Cajal body.

  • Authors: Song L, Han MH, Lesicka J, Fedoroff N
  • Journal: Proc Natl Acad Sci U S A 2007 13:5437-5442
  • PubMed ID: 17369351
Catalog #

Citations & References

Null mutation in macrophage migration inhibitory factor prevents muscle cell loss and fibrosis in partial bladder outlet obstruction.

  • Authors: Taylor JA, Zhu Q, Irwin B, Maghaydah Y, Tsimikas J, Pilbeam C, Leng L, Bucala R, Kuchel GA
  • Journal: Am J Physiol Renal Physiol 2006 6:F1343-F1353
  • PubMed ID: 16835407

Citations & References

IL-17 immunity in human type 1 diabetes.

  • Authors: Honkanen J, Nieminen JK, Gao R, Luopajarvi K, Salo HM, Ilonen J, Knip M, Otonkoski T, Vaarala O
  • Journal: J Immunol 2010 3:1959-1967
  • PubMed ID: 20592279

Citations & References

Comparison of gene expression profiles in mice liver following intravenous injection of 4 and 100 nm-sized PEG-coated gold nanoparticles

  • Authors: Cho, WS; Kim, S; Han, BS; Son, WC; Jeong, J
  • Journal: TOXICOLOGY LETTERS 2009 1:96-102
Catalog #

Citations & References

Alterations in the Noxa/Mcl-1 axis determine sensitivity of small cell lung cancer to the BH3 mimetic ABT-737.

  • Authors: Hauck P, Chao BH, Litz J, Krystal GW
  • Journal: Mol Cancer Ther 2009 4:883-892
  • PubMed ID: 19372561
Catalog #

Citations & References

Tissue factor-deficiency and protease activated receptor-1-deficiency reduce inflammation elicited by diet-induced steatohepatitis in mice.

  • Authors: Luyendyk JP, Sullivan BP, Guo GL, Wang R
  • Journal: Am J Pathol 2010 1:177-186
  • PubMed ID: 20008134

Citations & References

Histaminergic responses by hypothalamic neurons that regulate lordosis and their modulation by estradiol.

  • Authors: Dupre C, Lovett-Barron M, Pfaff DW, Kow LM
  • Journal: Proc Natl Acad Sci U S A 2010 27:12311-12316
  • PubMed ID: 20562342

Manual / Product Insert

MEGAshortscript™ Kit Protocol

Version: 1354M Rev E
Catalog #

Citations & References

Preliminary evidence for a modulation of fetal dopaminergic development by maternal immune activation during pregnancy

  • Authors: Meyer, U; Engler, A; Weber, L; Schedlowskic, M; Feldon, J
  • Journal: NEUROSCIENCE 2008 2:701-709
Catalog #

Citations & References

Myocardin inhibits cellular proliferation by inhibiting NF-kappaB(p65)-dependent cell cycle progression.

  • Authors: Tang RH, Zheng XL, Callis TE, Stansfield WE, He J, Baldwin AS, Wang DZ, Selzman CH
  • Journal: Proc Natl Acad Sci U S A 2008 9:3362-3367
  • PubMed ID: 18296632