Absolute Quantification of Specific Nucleic Acids by RT-PCR Using a Nonlinear Mathematical Model for Data Analysis

1994 ◽  
pp. 206-213
Keyword(s):  
Mathematical Model ◽  
Data Analysis ◽  
Nucleic Acids ◽  
Absolute Quantification ◽  
Rt Pcr ◽  
Nonlinear Mathematical Model

Detection of Nucleic Acids in Cells or Tissue Sections Using In situ Polymerase Chain Reaction (PCR)

1996 ◽  
Vol 54 ◽  
pp. 16-17
Author(s):  
J. R. Hully ◽  
K. R. Luehrsen ◽  
K. Aoyagi ◽  
C. Shoemaker ◽  
R. Abramson
Keyword(s):  
Nucleic Acids ◽  
Viral Infections ◽  
Anatomical Location ◽  
Rt Pcr ◽  
Reverse Transcription Pcr ◽  
Cellular Source ◽  
Gene Transcripts ◽  
Initial Description ◽  
In Cells

The development of PCR technology has greatly accelerated medical research at the genetic and molecular levels. Until recently, the inherent sensitivity of this technique has been limited to isolated preparations of nucleic acids which lack or at best have limited morphological information. With the obvious exception of cell lines, traditional PCR or reverse transcription-PCR (RT-PCR) cannot identify the cellular source of the amplified product. In contrast, in situ hybridization (ISH) by definition, defines the anatomical location of a gene and/or it’s product. However, this technique lacks the sensitivity of PCR and cannot routinely detect less than 10 to 20 copies per cell. Consequently, the localization of rare transcripts, latent viral infections, foreign or altered genes cannot be identified by this technique. In situ PCR or in situ RT-PCR is a combination of the two techniques, exploiting the sensitivity of PCR and the anatomical definition provided by ISH. Since it’s initial description considerable advances have been made in the application of in situ PCR, improvements in protocols, and the development of hardware dedicated to in situ PCR using conventional microscope slides. Our understanding of the importance of viral latency or viral burden in regards to HIV, HPV, and KSHV infections has benefited from this technique, enabling detection of single viral copies in cells or tissue otherwise thought to be normal. Clearly, this technique will be useful tool in pathobiology especially carcinogenesis, gene therapy and manipulations, the study of rare gene transcripts, and forensics.

scholarly journals Centrifugal step emulsification applied for absolute quantification of nucleic acids by digital droplet RPA

Lab on a Chip ◽  
2015 ◽  
Vol 15 (13) ◽  
pp. 2759-2766 ◽  
Author(s):  
Friedrich Schuler ◽  
Frank Schwemmer ◽  
Martin Trotter ◽  
Simon Wadle ◽  
Roland Zengerle ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Absolute Quantification ◽  
Recombinase Polymerase Amplification ◽  
Dead Volume

Centrifugal step emulsification enables dead volume free emulsification, it was applied to the first digital droplet recombinase polymerase amplification (RPA).

Quantitative real-time RT-PCR data analysis: current concepts and the novel “gene expression’s C T difference” formula

2006 ◽  
Vol 84 (11) ◽  
pp. 901-910 ◽  
Author(s):  
Jan H. Schefe ◽  
Kerstin E. Lehmann ◽  
Ivo R. Buschmann ◽  
Thomas Unger ◽  
Heiko Funke-Kaiser
Keyword(s):  
Data Analysis ◽  
Real Time ◽  
The Novel ◽  
Rt Pcr ◽  
Novel Gene ◽  
Difference Formula
Sign in / Sign up

Export Citation Format

Share Document