scholarly journals Successful gene expression analysis by multiplex, real-time, one-step RT-PCR, irrespective of the targets amplified

BioTechniques ◽  
2007 ◽  
Vol 43 (2) ◽  
pp. 230-231 ◽  
Author(s):  
Holger Engel ◽  
Corinna Kueppers ◽  
Margaretha Koenig ◽  
Dirk Loeffert
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Rt Pcr ◽  
One Step

scholarly journals Microdroplet-based one-step RT-PCR for ultrahigh throughput single-cell multiplex gene expression analysis and rare cell detection

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jennifer Ma ◽  
Gary Tran ◽  
Alwin M. D. Wan ◽  
Edmond W. K. Young ◽  
Eugenia Kumacheva ◽  
...  
Keyword(s):  
Gene Expression ◽  
Single Cell ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Single Cells ◽  
Cell Populations ◽  
Cell Detection ◽  
Rt Pcr ◽  
One Step ◽  
Cell Gene

AbstractGene expression analysis of individual cells enables characterization of heterogeneous and rare cell populations, yet widespread implementation of existing single-cell gene analysis techniques has been hindered due to limitations in scale, ease, and cost. Here, we present a novel microdroplet-based, one-step reverse-transcriptase polymerase chain reaction (RT-PCR) platform and demonstrate the detection of three targets simultaneously in over 100,000 single cells in a single experiment with a rapid read-out. Our customized reagent cocktail incorporates the bacteriophage T7 gene 2.5 protein to overcome cell lysate-mediated inhibition and allows for one-step RT-PCR of single cells encapsulated in nanoliter droplets. Fluorescent signals indicative of gene expressions are analyzed using a probabilistic deconvolution method to account for ambient RNA and cell doublets and produce single-cell gene signature profiles, as well as predict cell frequencies within heterogeneous samples. We also developed a simulation model to guide experimental design and optimize the accuracy and precision of the assay. Using mixtures of in vitro transcripts and murine cell lines, we demonstrated the detection of single RNA molecules and rare cell populations at a frequency of 0.1%. This low cost, sensitive, and adaptable technique will provide an accessible platform for high throughput single-cell analysis and enable a wide range of research and clinical applications.

Gene expression analysis of the rat testis after treatment with di(2-ethylhexyl) phthalate using cDNA microarray and real-time RT-PCR

2004 ◽  
Vol 200 (2) ◽  
pp. 103-110 ◽  
Author(s):  
Kazuyasu Kijima ◽  
Kaoru Toyosawa ◽  
Masashi Yasuba ◽  
Nobuo Matsuoka ◽  
Tetsuya Adachi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Expression Analysis ◽  
Cdna Microarray ◽  
Gene Expression Analysis ◽  
Rt Pcr ◽  
Rat Testis ◽  
Ethylhexyl Phthalate

scholarly journals Tissue-specific gene expression analysis of silkworm (Bombyx mori) by quantitative real-time RT-PCR

BMB Reports ◽  
2010 ◽  
Vol 43 (7) ◽  
pp. 480-484 ◽  
Author(s):  
Seung-Won Park ◽  
Seok-Woo Kang ◽  
Tae-Won Goo ◽  
Seong-Ryul Kim ◽  
Gwang-Gill Lee ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bombyx Mori ◽  
Real Time ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Specific Gene ◽  
Rt Pcr ◽  
Tissue Specific ◽  
Tissue Specific Gene ◽  
Silkworm Bombyx Mori

scholarly journals Microdissected Region-specific Gene Expression Analysis with Methacarn-fixed, Paraffin-embedded Tissues by Real-time RT-PCR

2004 ◽  
Vol 52 (7) ◽  
pp. 903-913 ◽  
Author(s):  
Hironori Takagi ◽  
Makoto Shibutani ◽  
Natsumi Kato ◽  
Haruka Fujita ◽  
Kyoung-Youl Lee ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Specific Gene ◽  
Rt Pcr ◽  
Specific Gene Expression
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