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Pyrosequencing is a way to measure methylation and genome variation in a relatively quick, high-throughput way. A region of DNA with either one or multiple CG (methylation) sites, or single nucleotide polymorphisms (SNPs) is sequenced in real time. The output is a percent methylation, or the percent of which allele is present in the SNP.
Pyrosequencing is a real-time sequence-synthesis based technology. Our machine, the Pyromark Q96 MD, can be used to quantify DNA methylation as well as for genotyping and SNP analysis for up to 96 samples per run. The machine detects the addition of each individual nucleotide as it is incorporated by generating light when a phosphate group is released.
See the video below for a demonstration of the pyro reaction cascade.
We help facilitate the study of epigenetic regulation mechanisms underlying normal development and disease pathogenesis.
Brunst KJ, Leung YK, Ryan PH, Khurana Hershey GK, Levin L, Ji H, Lemasters GK, Ho SM. Forkhead box protein 3 (FOXP3) hypermethylation is associated with diesel exhaust exposure and risk for childhood asthma. J Allergy Clin Immunol. 2013 Feb;131(2):592-4.e1-3. doi: 10.1016/j.jaci.2012.10.042. Epub 2012 Dec 20.
Lim E, Rothenberg ME. Demethylation of the human eotaxin-3 gene promoter leads to the elevated expression of eotaxin-3. J Immunol. 2014 Jan 1;192(1):466-74. doi 10.4049/jimmunol.1302454. Epub 2013 Dec 9.
Abdel-hameed, E.A., Ji H, Sherman KE, Shata MT. Epigenetic modification of FOXP3 in patients with chronic HIV infection. J Acquir Immune Defic Syndr. 2014 Jan 1;65(1):19-26. doi: 10.1097/QAI.0b013e3182a1bca4.
Zhang X, Ulm A, Somineni HK, Oh S, Weirauch MT, Zhang HX, Chen X, Lehn MA, Janssen EM, Ji H. DNA methylation dynamics during ex vivo differentiation and maturation of human dendritic cells. Epigenetics Chromatin. 2014 Aug 20;7:21. eCollection 2014.
Hong Ji, Xue Zhang, Sunghee Oh, Christopher N. Mayhew, Ashley Ulm, Hari K. Somineni, Mark Ericksen, James M. Wells, Gurjit K. Khurana Hershey. Dynamic transcriptional and epigenomic reprogramming from pediatric nasal epithelial cells to induced pluripotent stem cells. JACI, Oct, 2014. DOI: http://dx.doi.org/10.1016/j.jaci.2014.08.038.
Expressions of methyltransferase enzymes correlate with FOXP3 methylation in the colon mucosa of HIV-1 infected patients. Abdel-hameed, E.A., Ji, H., Sherman, K., Shata, MT. (2012) 14th Annual Meeting of the Institute of Human Virology (October 14-17, 2012 in Baltimore, Maryland).
The Role of Epigenetic Modification of FOXP3 in Regulation of the Immune Response in HIV/HCV infection. Abdel-hameed, E.A., Ji, H., Sherman, K., Shata, MT. (2012) Digestive Health Center Annual Scientific Retreat, CCHMC February 28, 2012.
Identification of Epigenetic Variations associated with Air Pollution and Asthma. Noelle Jurcak, Ashley Ulm, Hari K. Somineni, Xue Zhang, Sharon Sauter, Joceyln Biagini Myers, Melinda Butsch Kovacic, and Hong Ji. SURF. 2014 Aug.
Genetic variation in the FADS gene family and Docosahexaenoic Acid (DHA) Levels in Randomized trial of DHA- Supplemented Mothers. Christina J. Valentine, Alexa M. Anderson, Kelly Dingess, Jeanne Kleiman, Ardythe Morrow, and Lynette Rogers. Cincinnati Children’s Hospital, August 1 2014.
Pyrosequencing Core Director:Hong Ji, PhD Phone: 513-803-5055
Pyrosequencing Core Manager:Ashley Ulm Phone: 513-803-0562
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