In Situ PCR

The In Situ PCR:
(Amplification and Detection in a Cellular Context)

Ernest F. Retzel, Katherine A. Staskus, Janet E. Embretson and Ashley T. Haase
Department of Microbiology, University of Minnesota, Minneapolis, MN 55455

Table of Contents
Introduction Technical Aspects/Philosophy Primer Design Reaction Conditions "Mechanics" of the Reaction Methodology
A. Cell and Tissues.
1. Cultured Cells: Infection and Preparation. 2. Tissues: Pulmonary infection and preparation.
B. In Situ Amplification
1. Cultured Cells in Suspension. 2. Tissues on Slides.
C. Detection of Amplified DNA by in situ Hybridization
1. Solution amplification. 2. Tissue Sections.
References

In Situ PCR on Plant Material (Bo Johansen)Required materialsRequired solutionsTissue preparationPectinase treatmentProtease treatmentAcetylationDNase treatmentReverse transcriptionThermal cyclingDetectionMountingAcrobat version of PosterReferencesOther ISPCR linksBotanical Institute

RT IN SITU PCR
Gerard J. Nuovo
TABLE OF CONTENTS
1. Abstract
2. Introductory statement
3. The key preparatory steps
3.1. Fixative
3.1.1. Protease digestion
3.1.2. Definition of optimal protease digestion
3.1.3. Definitions of suboptimal and over-digestion
3.1.4. DNase digestion
3.1.5. Direct incorporation of the reporter nucleotide
4. A one step protocol for RT in situ PCR
5. Applications of RT in situ PCR Matrix metalloprotease expression in cancer as a model for RT in situ PCR
5.1. MMP and TIMP expression in cervical cancer
6. Concluding remarks
7. References
8. Entire manuscript

In situ PCR: protocols and applications.
groups have published protocols and data using in situ PCR techniques. (5). This article will discuss some of the basic concepts of in situ PCR, the proto- ...www.genome.org/cgi/reprint/4/4/S151.pdf

Co-labeling Using In Situ PCR: A Review
Figure 2 Background vs signal with RT in situ PCR. This renal biopsy was analyzed for mRNA by RT in situ PCR. After digestion for 10 min in ...www.jhc.org/cgi/reprint/49/11/1329.pdf

Protocols for the : in situ: PCR-amplification and detection of mRNA and DNA sequencing.
In this protocol we describe the in situ PCR method for the amplification of both DNA and mRNA targets [in situ reverse transcriptase-PCR (RT-PCR)], ...www.nature.com/nprot/journal/v2/n11/abs/nprot.2007.395.html

96-well RNA In Situ Hybridization Protocol
Materials. 1.1. Probe Preparation. 1.1.1. Cell Inoculation. 1. 96-well, deep square-well round bottom plate (E&K Scientific Ritter Riplate). ...www.fruitfly.org/about/methods/RNAinsitu.html

A protocol for PCR in situ hybridization of hyphomycetesIntroduction. Within the last years the focus of mycological studies has shifted. emphasis from mere taxonomic studies to more ecological. questions. ...www.im.microbios.org/03setember98/08%20Sterflinger.pdf