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IMolecular Biology / PCR / Variants of PCR / 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 materials
Required solutions
Tissue preparation
Pectinase treatment
Protease treatment
Acetylation
DNase treatment
Reverse transcription
Thermal cycling
Detection
Mounting
Acrobat version of Poster
References
Other ISPCR links
Botanical 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.htmlw.

 

96-well RNA In Situ Hybridization Protocol

  1. 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 hyphomycetes
Introduction. 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

 

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