The use of reverse transcription polymerase chain reaction to analyse large numbers of mRNA species from a single cell

K M Toellner; D Scheel-Toellner; U Seitzer; R Sprenger; L Trümper; C Schlüter; H D Flad; J Gerdes

The use of reverse transcription polymerase chain reaction to analyse large numbers of mRNA species from a single cell

K M Toellner, D Scheel-Toellner, U Seitzer, R Sprenger, L Trümper, C Schlüter, H D Flad, J Gerdes

Immunology and Immunotherapy

Research output: Contribution to journal › Article › peer-review

19 Citations (Scopus)

Abstract

A PCR method is described for determining the expression of multiple heterogeneous mRNAs from single cells. The total mRNA pool of a single selected cell is subjected to reverse transcription and subsequent tailing with poly(dA). This cDNA is preamplified by a sequence non-specific PCR protocol using oligo(dT)-containing primers. The single cell cDNA library obtained permits the analysis of virtually unlimited numbers of mRNA species per cell using sequence-specific PCR. This procedure of multiple mRNA analysis enables phenotyping of any cell for its mRNA composition and could be used to study the cytokine mRNA expression of individual human T cells ex vivo. The method should greatly facilitate the analysis of combinatorial expression of known genes in any cell.

Original language	English
Pages (from-to)	71-5
Number of pages	5
Journal	Journal of Immunological Methods
Volume	191
Issue number	1
Publication status	Published - 10 May 1996

Keywords

RNA, Messenger
Polymerase Chain Reaction
DNA, Complementary
Base Sequence
Leukocytes, Mononuclear
Humans
Molecular Sequence Data
Transcription, Genetic

Cite this

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abstract = "A PCR method is described for determining the expression of multiple heterogeneous mRNAs from single cells. The total mRNA pool of a single selected cell is subjected to reverse transcription and subsequent tailing with poly(dA). This cDNA is preamplified by a sequence non-specific PCR protocol using oligo(dT)-containing primers. The single cell cDNA library obtained permits the analysis of virtually unlimited numbers of mRNA species per cell using sequence-specific PCR. This procedure of multiple mRNA analysis enables phenotyping of any cell for its mRNA composition and could be used to study the cytokine mRNA expression of individual human T cells ex vivo. The method should greatly facilitate the analysis of combinatorial expression of known genes in any cell.",

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T1 - The use of reverse transcription polymerase chain reaction to analyse large numbers of mRNA species from a single cell

AU - Toellner, K M

AU - Scheel-Toellner, D

AU - Seitzer, U

AU - Sprenger, R

AU - Trümper, L

AU - Schlüter, C

AU - Flad, H D

AU - Gerdes, J

PY - 1996/5/10

Y1 - 1996/5/10

N2 - A PCR method is described for determining the expression of multiple heterogeneous mRNAs from single cells. The total mRNA pool of a single selected cell is subjected to reverse transcription and subsequent tailing with poly(dA). This cDNA is preamplified by a sequence non-specific PCR protocol using oligo(dT)-containing primers. The single cell cDNA library obtained permits the analysis of virtually unlimited numbers of mRNA species per cell using sequence-specific PCR. This procedure of multiple mRNA analysis enables phenotyping of any cell for its mRNA composition and could be used to study the cytokine mRNA expression of individual human T cells ex vivo. The method should greatly facilitate the analysis of combinatorial expression of known genes in any cell.

AB - A PCR method is described for determining the expression of multiple heterogeneous mRNAs from single cells. The total mRNA pool of a single selected cell is subjected to reverse transcription and subsequent tailing with poly(dA). This cDNA is preamplified by a sequence non-specific PCR protocol using oligo(dT)-containing primers. The single cell cDNA library obtained permits the analysis of virtually unlimited numbers of mRNA species per cell using sequence-specific PCR. This procedure of multiple mRNA analysis enables phenotyping of any cell for its mRNA composition and could be used to study the cytokine mRNA expression of individual human T cells ex vivo. The method should greatly facilitate the analysis of combinatorial expression of known genes in any cell.

KW - RNA, Messenger

KW - Polymerase Chain Reaction

KW - DNA, Complementary

KW - Base Sequence

KW - Leukocytes, Mononuclear

KW - Humans

KW - Molecular Sequence Data

KW - Transcription, Genetic

M3 - Article

C2 - 8642203

SN - 0022-1759

VL - 191

SP - 71

EP - 75

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

IS - 1

ER -

The use of reverse transcription polymerase chain reaction to analyse large numbers of mRNA species from a single cell

Abstract

Keywords

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