Bassine Science Building, 103
DegreesRockefeller University, Ph.D.
Brandeis University, B.A.
ExpertiseLinear-After-The-Exponential (LATE)-PCR technique of DNA amplification and it's application to diagnostic assays.
ProfileScientists in our laboratory have overcome the inherent limitations of symmetric PCR by developing an advanced form of asymmetric PCR, known as LATE-PCR. Symmetric PCR has been the dominant method of DNA amplification for the last 20 years. Each amplicon in such assays is generated by use of two primers of equal concentration and equal Tm (melting temperature) and is therefore double-stranded. Amplification proceeds exponentially, but then slows down and reaches plateau in a stochastic manner. Thus, although such reactions are well known and are often robust, they nevertheless suffer from limitations which are particularly evident in samples containing low numbers of initial targets. These limitations include:
* Design difficulties, particularly for multiplexing
* Reduced efficiency and sensitivity due to mis-priming
* Variability among replicate reactions, particularly at end-point
* Limited availability of fluorescent colors
* Limitations in probe design
* Requirement for hot-start enzymes
|BIOL||350a||Graduate Student Research Seminar|
|BIOL||350b||Graduate Student Research Seminar|
|BIOL||350d||Graduate Student Research Seminar|
|BISC||2b||Genes and the Human Story|
Awards and Honors
2009-2011 Sigma Xi Society - Distinguished Lecturer (2009)
Feldman Brothers Award for Course Development in human reproduction and population explosion (1999)
Helen Hay Whitney Postdoctoral Fellowship (1973 - 1975)
Harald Peter, Grace Henihan, Kenneth E. Pierce, Kate Templeton, Rohit Mistry, Carmelo Volpe, Lawrence J. Wangh, Till T. Bachmann. "Taking the Bull by the Horns – A new High Content Molecular Assay Addressing Complex ESBL Detection." European Society of Clinical Microbiology and Infectious Disease, March 2012. London, England
John E. Rice, Arthur H. Reis, Jr., Lisa M. Rice, Rachel K. Carver-Brown, Shana Augustin Tetrault & Lawrence J. Wangh. "Fluorescent Signatures for Variable DNA Sequences." Nucleic Acid Research NAR 1-10, doi:10.1093/nar/gks731 (Featured Article). (2012).
Kenneth E. Pierce and Lawrence J. Wangh. "Rapid detection and sequence analysis of Hepatitis C Virus using a novel molecular assay with mismatch-tolerant hybridization probes." Journal of Clinical Microbiology manuscript in preparation. (2012).
Kenneth E. Pierce and Lawrence J. Wangh. "Rapid detection and sequence analysis of Hepatitis C Virus using a novel molecular assay with mismatch-tolerant hybridization probes." CFAR. Harvard Medical School. April 2012.
Kenneth E. Pierce, Harald Peter, Till T. Bachman, Carmelo Volpe, John E. Rice, and Lawrence J. Wangh. "Rapid Detection of TEM-type Extended Spectrum Beta Lactamase (ESBL) mutations using multiple Lights-On-Lights-Off Probes with single-stranded DNA amplification." Journal of Applied Microbiology SCO 5.1.0 DTD JMDI219_proof 16 March 2013 8:00 pm EO: JMD12_0148. (2012).
Kenneth E. Pierce, Huma Khan, Rohit Mistry, Simon D. Goldenberg, Gary L. French, and Lawrence J. Wangh. "RAPID DETECTION OF SEQUENCE VARIATION IN CLOSTRIUM DIFFICILE GENES USING LATE-PCR WITH MULTIPLE MISMATCH-TOLERANT HYBRIDIZATION PROBES." Journal of Clinical Microbiology Journal of Microbiological Methods, 91 (2012) 269–275.. (2012).
L. Rice, J. Rice, S. Fishbein, L. Wangh, N. Kurpina, B. Kreiswirth, N. Casali, F. Drobniewski, EM. Streicher, and R. Warren. "Construction and Evaluation of an M(X)DR-TB Multiplex Assay using LATE-PCR & Thermalight™ Probes." 2012 New England TB Symposium. Broad Institute, Cambridge MA. June 29, 2012.
L. Wangh, J. Rice, L. Rahman, B. Ronish, S. Fishbein, N. Kurpina, B. Kreiswirth, N. Casali, F. Drobniewski, EM. Streicher, and R. Warren. "Evaluation of an M(X)DR-TB Multiplex Assay using LATE-PCR & Thermalight™ Probes." Keystone Symposium: Drug Resistance and Persistance in Tuberculosis. Kampala Uganda. May 13-18, 2012.
L. Wangh, J. Rice, L. Rice, S. Fishbein, L. Rahman, B. Ronish, N. Kurpina, B. Kreiswirth N. Casali, F. Drobniewski, and Rohit Mistry. "Two Single-Tube Multiplex Assays for M(X)DR-TB and NTMs using LATE-PCR & Thermalight™ Probes." World TB Day 2012. Boston University Medical School. March 23, 2012.
Lisa M. Rice, Arthur H. Reis, Jr., Bonnie Ronish, Rachel K. Carver-Brown, Greg Williams, John Czajka, Nicole Gentile, Gerald Kost, and Lawrence Wangh. "Design of a Single-Tube, Endpoint, LATE-PCR Assay for Seventeen Pathogens Associated with Sepsis." Journal of Applied Microbiology manuscript submitted. (2012).
Nicola Casali, Agnieszka Broda, John Rice, Rohit Mistry, Carmelo Volpe, Larry Wangh, Jay Lewington and Francis Drobniewski. "A Novel Single-Tube PCR Assay for Rapid Identification of MDR-TB." European Society of Clinical Microbiology and Infectious Diseases, March 2012. London, England
Nicole L. Gentile, Anna M. Dillier, Greg V. Williams, Jared Ackers, Arthur H. Reis, Jr., Lisa M. Rice, Lawrence Wangh, John Czajka, Gerald J. Kost. "Validation of Monoplex and Multiplex LATE-PCR Gene Specific Sepsis Assays Using Clinical Isolates." Journal of Applied Microbiology manuscript submitted. (2012).
Osborne, Adam E., Wangh, Lawrence J.. "A LATE-PCR Decaplex Assay for the Detection and Sequencing of Mitochondrial DNA Mutations Down to the Single Genome Level." PloS ONE manuscript submitted. (2012).
Rachel K. Carver-Brown, Arthur H. Reis, Jr., Lisa Rice, Nicole Gentile, Wanvisa Boonlert, Gerald Kost, and Lawrence Wangh. "A Single Tube LATE-PCR Multiplex Assay with Lights-On/Lights-Off Probes for the Detection of Septicemia." Journal of Applied Microbiology manuscript in preparation. (2012).
The Wangh Laboratory Site. LATE-PCR.Org. Brandeis University
Wangh, Lawrence J. "Advances in LATE-PCR and Its Allied Technologies." Select BioSciences: Genomics Research - Advances in qPCR. April 19-20, Boston. http://selectbiosciences.com/conferences/agenda.aspx?pid=190&conf=AqPCR2012.
Wangh, Lawrence J. "Advantages of LATE-PCR Technology." (2012): <http://www.bioquicknews.com/node/761>.
Wangh, Lawrence J. "Very Highly Informative Single-Tube Assays for M(X)DR-TB and Non-Tuberculosis Mycobacteria." Select Biosciences: Molecular Diagnostics Europe, Point of Care Diagnostics, May 10-11, 2012. London, England. http://selectbiosciences.com/conferences/agenda.aspx?pid=350&conf=POC2012.
Wangh, Lawrence J. "GEN Tech Tips PCR: Trends and New Directions." (2011): <http://www.genengnews.com/gen-articles/pcr-trends-and-new-directions/3946/>.
Wangh, Lawrence J. "Genetic Engineering News: “Scaling PCR Workflows from Benchtop to Automation.” Webinar, 29 June 2010." (2011)
Wangh, Lawrence J. "Genome Web: http://www.genomeweb.com/dxpgx/smiths-detection-inks-mdx-license-novartis." (2011)
B. Ronish, M. Hakhverdyan, K. Ståhl, S. Belák, N. LeBlanc, L. Wangh. "Design and Verification of a highly reliable Linear-After-The-Exponential PCR assay for the detection of African Swine Fever virus." J. Veterinary Methods http://dx.doi.org/10.1016/j.jviromet.2010.12.003 (2010).
Hartshorn, Cristina, and Wangh, Lawrence J. "One-step RT-LATE-PCR for mRNA and viral RNA detection and quantification. RT-PCR Protocols: Second Edition." Methods in Molecular Biology series. vol. 2 New York, NY: Springer, 2010
Jia, Y., Hartshorn, C., Hartung, O, Wangh, L.J.. "Heat Shock Memory in Preimplantation Mouse Embryos,." . Fertility and Sterility Vol. 93, No. 8, May 15, 2010 (2010).
Jia, Y., Osborne, A., Rice, J.E., and Wangh, L.J.. "Dilute-‘N’-Go Dideoxy Sequencing of All DNA Strands Generated in Multiplex LATE-PCR Assays." Nucleic Acids Research, doi: 10.1093/nar/gkq111 (2010).
Pierce, K., Mistry, R., Reid, S.M., Bharya, S., Dukes, J.P., Hartshorn, C., King., D.P., Wangh, Lawrence J. "Design and optimization of a novel reverse transcription linear-after-the-exponential PCR for the detection of foot-and-mouth disease virus.." Journal of Applied Microbiology doi:10.1111/j.1365-2672.2009.04640.x online. (2010).
Osborne, A., Reis, A., Bach, L., Wangh, L.. "Single-Molecule LATE-PCR Analysis of Human Mitochondrial Genomic Sequence Variations." PLoS One 4(5). (2009).
Hartshorn, C., Anshelevich, A., Jia, Yanwei, and Wangh, L.J.. "Early Onset of Heat-Shock Response in Mouse Embryos Revealed by Quantification of Stress-Inducible hsp70i RNA." Gene Regulation and Systems Biology 1. (2007): 1-9.
Hartshorn, C., Eckert, J.J., Hartung, O., Wangh, L.J.. "Oct4 and Xist RNA gradients in 8-cell embryos revealed by single-cell duplex RT-LATE-PCR." BMC Biotechnology, 7. 87 (2007): doi:10.1186/1472-6750-7-87.
John Rice, J. Aquiles Sanchez, Kenneth E. Pierce, Arthur Reis, and Lawrence J. Wangh. "LATE-PCR Multi-plexing using PrimeSafe® followed by Multi-Dilute-`N'-Go Dideoxy Sequencing." Nature Protocols 2. 10 (2007): 2429-2438..
Pierce, K.E. and Wangh, L.J., ed. Linear-After-The-Exponential Polymerase Chain Reaction and Allied Technologies:. Methods and Protocols, 2007.
Wangh, Lawrence J. Podcast on LATE-PCR. February 22, 2007. Genetic Engineering News,. <http://www.genengnews.com/genCasts.aspx?id=79>.
J. Aquiles Sanchez, Jessica Abramovitz, Jesse J. Salk, Arthur H Reis, John E Rice, Kenneth E Pierce, and Lawrence J. Wangh. "Two-temperature LATE-PCR endpoint genotyping." BMC Biotechnology 6. 44 (2006).
Jesse J. Salk, J Aquiles Sanchez, Kenneth E. Pierce, John E. Rice, Kevin C. Soares, Lawrence J. Wangh. "Direct Amplification of Single-Stranded DNA for Pyrosequencing using Linear-After-The-Exponential (LATE)-PCR." Analytical Biochemistry Volume 353. Issue 1 (2006): 124-132.
Kenneth E. Pierce and Lawrence J. Wangh, Editor: Alan Thornhill /Series Editor: John Walker, ed. LATE-PCR and allied technologies: Real-time detection strategies for rapid, reliable diagnosis from single cells. Humana Press: UK, 2006.
http://www.brandeis.edu/wanghlab/. Website for LATE-PCR. March 2007. Brandeis University. <http://www.brandeis.edu/wanghlab/>.
Hartshorn, C., Anshelevich, A., and Wangh, L.J.. "Laser zona-drilling does not induce hsp70i transcription in blastomeres of 8-cell mouse embryos." Fertility and Sterility 84. (5) (2005): 1547-1550.
Hartshorn, C., Anshelevich, A., and Wangh, L.J.. "Rapid, single-tube method for quantitative preparation and analysis of RNA and DNA in samples as small as one cell." BMC Biotechnology 5. (2005): 2.
Pierce, K.E., Sanchez, J.A., Rice, J.E., and Wangh, L.J. "Linear-After-The-Exponential (LATE)-PCR: Primer design criteria for high yields of specific single-stranded DNA and improved real-time detection." PNAS 102:. (2005): 8609-8614.
Pierce, K.E., and Wangh, L.J.. "Effectiveness and limitations of uracil-DNA glycosylases in sensitive real-time PCR assays." BioTechniques 36. (1) (2004): 44-48.
Sanchez, J.A., Pierce, K.E., Rice, J.E., and Wangh, L.J. "Linear-After-The-Exponential (LATE)-PCR: An advanced method of asymmetric PCR and its uses in quantitative real-time analysis." PNAS 101. (7) (2004): 1933-1938.
Hartshorn C, Rice JE, Wangh LJ.. "Differential pattern of Xist RNA accumulation in single blastomeres isolated from 8-cell stage mouse embryos following laser zona drilling." Molecular Reproduction and Development 64. (2003): 41-51.
Pierce, K.E., Rice, J.E., Sanchez, J.A., and Wangh, L.J.. "Detection of cystic fibrosis alleles from single cells using molecular beacons and a novel method of asymmetric real-time PCR." Molecular Human Reproduction 9. (12) (2003): 815-820.
Hartshorn, C., Rice, J.E., and Wangh, L.J.. "Developmentally-Regulated Changes of Xist RNA Levels in Single Preimplantation Mouse Embryos, as Revealed by Quantitative Real-Time PCR." Molecular Reproduction and Development 61. (2002): 425-436.
Pierce, K., Rice, J.E., Sanchez, J.A., and Wangh, L.J.. "QuantiLyse: Reliable DNA amplification from single cells." BioTechniques 32. (5) (2002): 1106-1111.
Rice, J.E., Sanchez, J.A., Pierce, K.E., and Wangh, L.J.. "Real-time PCR with molecular beacons provides a highly accurate assay for detection of Tay-Sachs alleles in single cells." Prenatal Diagnosis 22. (2002): 113-1134.
Pierce, K.E., Rice, J.E., Sanchez, J.A., Brenner, C., and Wangh, L.J.. "Real-time PCR using molecular beacons for accurate detection of the Y chromosome in single human blastomeres." Molecular Human Genetics 6. (12) (2000): 1155-1164.