Description(s):

Gene expression is regulated by individual cells, so why not study gene expression in a single cell such as stem cells, neurons, developmental tissues, and laser capture microsections? To allow researchers the ability to take on single cell studies, Signosis has developed a single cell SYBR Green based real-time RT-PCR Assay kit which allows you to directly assay gene expression levels in a single cell using cell lysate without any RNA preparation. This kit provides a simple, sensitive, quantitative and cost-effective method to monitor PCR reactions in real time. The reagents for cDNA synthesis and PCR amplification in addition to Direct cDNA cell lysis buffer are also included in the kit.

Benefits:

All-in-One - All reagents for the experiment, from cell lysis, reverse transcription to PCR assay, are included in  one kit. No need for RNA preparation - Cell lysates can be used directly in reverse transcription. Robust - Full length cDNA can be obtained with the kit. Real-time - Analysis can be monitored in real time.

Principle

The buffers are optimized for cell lysate preparation and real-time RT-PCR.

 

Data

Detection of Limited Target Sequences from Cell Lysate. Increasing amounts of HelA cells were lysed with Direct cDNA cell lysis buffer. All samples were reverse transcribed by the RT reagents provided in the kit. Real-time PCR was conducted for beta-actin, GAPDH and B2M genes on all samples in triplicate reactions on a 7700 system.

 

Citations

 

1.Inositol 1,4,5-trisphosphate (IP3) receptor up-regulation in hypertension is associated with sensitization of Ca2+ release and vascular smooth muscle contractility. Abou-Saleh H, Pathan AR, Daalis A, Hubrack S, Abou-Jassoum H, Al-Naeimi H, Rusch NJ, Machaca K J Biol Chem. 2013 41958;288(46):32941-51. 

2.A long noncoding RNA contributes to neuropathic pain by silencing Kcna2 in primary afferent neurons. Zhao X, Tang Z, Zhang H, Atianjoh FE, Zhao JY, Liang L, Wang W, Guan X, Kao SC, Tiwari V, Gao YJ, Hoffman PN, Cui H, Li M, Dong X, Tao YX. Nat Neurosci. 2013 Aug;16(8):1024-31. 

3.Protein Kinase CK2 Inhibition Down Modulates the NF-κB and STAT3 Survival Pathways, Enhances the Cellular Proteotoxic Stress and Synergistically Boosts the Cytotoxic Effect of Bortezomib on Multiple Myeloma and Mantle Cell Lymphoma Cells. Manni S, Brancalion A, Mandato E, Tubi LQ, Colpo A, Pizzi M, Cappellesso R, Zaffino F, Di Maggio SA, Cabrelle A, Marino F, Zambello R, Trentin L, Adami F, Gurrieri C, Semenzato G, Piazza F. PLoS One.  2013 41544;8(9):e75280. 

4.IP3 receptor up-regulation in hypertension is associated with sensitization of Ca2+ release and vascular smooth muscle contractility. Abou-Saleh H, Pathan AR, Daalis A, Hubrack S, Abou-Jassoum H, Al-Naeimi H, Rusch NJ, Machaca K. J Biol Chem.  2013 Oct 4. [Epub ahead of print] 

5.Direct Quantification of mRNA and miRNA from Cell Lysates Using Reverse Transcription Real Time PCR: A Multidimensional Analysis of the Performance of Reagents and Workflows. Ho YK, Xu WT, Too HP. PLoS One.  2013 41522;8(9):e72463. 

6.Osteoprotegerin Contributes to the Metastatic Potential of Cells with a Dysfunctional TSC2 Tumor-Suppressor Gene. Steagall WK, Pacheco-Rodriguez G, Glasgow CG, Ikeda Y, Lin JP, Zheng G, Moss J. Am J Pathol.  2013 Sep;183(3):938-50. 

7.Rapid expression profiling of brain microvascular endothelial cells by immuno-laser capture microdissection coupled to TaqMan® Low Density Array. Demarest TG, Murugesan N, Shrestha B, Pachter JS. J Neurosci Methods., 2012 ;206(2):200-4. 

8.Nuclear Aggregation of Olfactory Receptor Genes Governs Their Monogenic Expression.Clowney EJ, Legros MA, Mosley CP, Clowney FG, Markenskoff-Papadimitriou EC, Myllys M, Barnea G, Larabell CA, Lomvardas S. Cell., 2012 41222;151(4):724-37. 

9.Active induction of experimental autoimmune encephalomyelitis by MOG35-55 peptide immunization is associated with differential responses in separate compartments of the choroid plexus. Murugesan N, Paul D, Lemire Y, Shrestha B, Ge S, Pachter JS. Fluids Barriers CNS., 2012 41125;9(1):15. 

10. Alterations in Tight Junction Protein and IgG Permeability Accompany Leukocyte Extravasation Across the Choroid Plexus During Neuroinflammation.Shrestha, Bandana BS; Paul, Debayon MS; Pachter, Joel S. PhD . Journal of neuropathology and experimental …, 2014

11. Chronological in Vivo Imaging Reveals Endothelial Inflammation Prior to Neutrophils Accumulation and Lipid Deposition in HCD-Fed Zebrafish. Atherosclerosis. Luo, Hui, et al.  2019, doi:10.1016/j.atherosclerosis.2019.09.017.

12. N6‐Methyladenosine Demethylase FTO Contributes to Neuropathic Pain by Stabilizing G9a Expression in Primary Sensory Neurons. Li, Y., Guo, X., Sun, L., Xiao, J., Su, S., Du, S., Li, Z., Wu, S., Liu, W., Mo, K., Xia, S., Chang, Y.‐J., Denis, D. and Tao, Y.‐X. (2020). Adv. Sci.. doi:10.1002/advs.201902402

13. The transcription factor TAL1 and miR-17-92 create a regulatory loop in hematopoiesis. Meyer, A., Herkt, S., Kunze-Schumacher, H. et al. Sci Rep 10, 21438 (2020). https://doi.org/10.1038/s41598-020-78629-z

14. Downregulation of a Dorsal Root Ganglion-Specifically Enriched Long Noncoding RNA is Required for Neuropathic Pain by Negatively Regulating RALY-Triggered Ehmt2 Expression. Pan, Z., Du, S., Wang, K., Guo, X., Mao, Q., Feng, X., Huang, L., Wu, S., Hou, B., Chang, Y.‐J., Liu, T., Chen, T., Li, H., Bachmann, T., Bekker, A., Hu, H., Tao, Y.‐X. Adv. Sci. 2021, 2004515. https://doi.org/10.1002/advs.202004515