Absolute quantification of mrna using real-time reverse transcription polymerase chain reaction assays

Journal of Molecular Endocrinology

Volumn 25, Issue 2, 2000, Pages 169-193

  Bustin, S A ^a  

^a QUEEN MARY UNIVERSITY OF LONDON   (United Kingdom)

Author keywords

[No Author keywords available]

Indexed keywords

ACCURACY; FLUORESCENCE; GENE EXPRESSION; GENETIC TRANSCRIPTION; HOUSEKEEPING GENE; PRIORITY JOURNAL; REPRODUCIBILITY; REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION; REVIEW;

EID: 0033775615     PISSN: 09525041     EISSN: None     Source Type: Journal    
DOI: 10.1677/jme.0.0250169     Document Type: Review

References (216)

1. Thermostability and thermoactivity of enzymes from hyperthermophilic Archaea
2. Use of two reporter dyes without interference in a single-tube rapidcycle PCR: Alpha(1)-antitrypsin genotyping by multiplex real-time fluorescence PCR with the LightCycler
3. Insulin stimulates glyceraldehyde-3-phosphate dehydrogenase gene expression through cis-acting DNA sequences
4. Multiple insulin-responsive elements regulate transcription of the GAPDH gene
5. Rapid detection of deletion mutations in inherited metabolic diseases by melting curve analysis with LightCycler
6. Identification of bone morphogenetic proteins and their receptors in human breast cancer cell lines: Importance of bmp2
7. High-speed prothrombin G A 20210 and methylenetetrahydrofolate reductase C T 677 mutation detection using real-time fluorescence PCR and melting curves
8. Northern blot normalization with a 28S rRNA oligonucleotide probe
9. Norepinephrine, tri-iodothyronine and insulin upregulate glyceraldehyde-3-phosphate dehydrogenase mRNA during Brown adipocyte differentiation
10. Use of manganese in RT-PCR eliminates PCR artifacts resulting from DNase I digestion
11. Absolute levels of mRNA by polymerase chain reaction-aided transcript titration assay
12. On the fidelity of DNA replication: Manganese mutagenesis in vitro
13. Quantification of viral load: Clinical relevance for human immunodeficiency virus, hepatitis B virus and hepatitis C virus infection
14. Homogeneous multiplex genotyping of hemochromatosis mutations with fluorescent hybridization probes
15. Integrated amplification and detection of the C677T point mutation in the methylenetetrahydrofolate reductase gene by fluorescence resonance energy transfer and probe melting curves
16. Color multiplexing hybridization probes using the apolipoprotein E locus as a model system for genotyping
17. Comparison of glyceraldehyde-3-phosphate dehydrogenase and 28S-ribosomal RNA gene expression as RNA loading controls for northern blot analysis of cell lines of varying malignant potential
18. Quantitation of MYC gene expression in sporadic breast tumors with a real-time reverse transcription-PCR assay
19. Real-time reverse transcription-PCR assay for future management of ERBB2-based clinical applications
20. Quantitative reverse transcriptase-PCR amplification of cytokine mRNA in liver biopsy specimens using a non-competitive method
21. LightCycler PCR assay for simultaneous detection of the H63D and S65C mutations in the HFE hemochromatosis gene based on opposite melting temperature shifts
22. Thermodynamic basis of the enhanced specificity of structured DNA probes
23. A lock-docking oligo(dT) primer for 5' and 3' RACE PCR
24. Direct quantitation of RNA transcripts by competitive single-tube RT-PCR and capillary electrophoresis
25. Secondary structure in the 3' UTR of EGF and the choice of reverse transcriptases affect the detection of message diversity by RT-PCR
26. Regulatory elements and expression profiles
27. Synthesis of full length cDNAs from four partially purified oviduct mRNAs
28. Molecular assessment of tumour stage and disease recurrence using PCR-based assays
29. The product of the primary response gene BRF1 inhibits the interaction between 14-13-3 proteins and cRaf-1 in the yeast trihybrid system
30. Detection of cytokeratins 19/20 and guanylyl cyclase C in peripheral blood of colorectal cancer patients
31. Pregnancy induces an increase in the expression of glyceraldehyde-3-phosphate dehydrogenase in uterine artery endothelial cells
32. Pancreatic GAPDH gene expression during ontogeny and acute pancreatitis induced by caerulein
33. Use of real-time PCR and fluorimetry to detect lamivudine resistance-associated mutations in hepatitis B virus
34. Changes in beta-actin mRNA expression in remodeling canine myocardium
35. Up-regulation of beta-actin, cyclophilin and GAPDH in N1S1 rat hepatoma
36. Coordinated induction of two unrelated glucose-regulated protein genes by a calcium ionophore: Human BiP/GRP78 and GAPDH
37. Illegitimate transcription: Transcription of any gene in any cell type
38. Involvement of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and p53 in neuronal apoptosis: Evidence that GAPDH is upregulated by p53
39. Alternative splicing of mRNA as a mode of endocrine regulation
40. Highly sensitive method to detect mRNAs in individual cells by direct RT-PCR using Tth DNA polymerase
41. PCR fidelity of pfu DNA polymerase and other thermostable DNA polymerases
42. Peptide nucleic acids: Expanding the scope of nucleic acid recognition
43. Improved reverse transcriptase-polymerase chain reaction protocol with exogenous internal competitive control for prostate-specific antigen mRNA in blood and bone marrow
44. Comparison of M-MLV reverse transcriptase and Tth polymerase activity in RT-PCR of samples with low virus burden
45. Amplification of human beta-actin gene by the reverse transcriptase-polymerase chain reaction: Implications for assessment of RNA from formalin-fixed, paraffin-embedded material
46. Measurement of sputum Mycobacterium tuberculosis messenger RNA as a surrogate for response to chemotherapy
47. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, E.C. 1.2.1.12.) gene expression in two malignant human mammary epithelial cell lines: BT-20 and MCF-7. Regulation of gene expression by 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3)
48. Polymerization and RNase H activities of the reverse transcriptases from avian myeloblastosis, human immunodeficiency, and Moloney murine leukemia viruses are functionally uncoupled
49. Detection of cytokeratin-19 transcripts by reverse transcriptase-polymerase chain reaction in lung cancer cell lines and blood of lung cancer patients
50. Human beta-actin retropseudogenes interfere with RT-PCR
51. ASDB: Database of alternatively spliced genes
52. DNA polymerase fidelity and the polymerase chain reaction
53. Potential of LightCycler technology for quantification of minimal residual disease in childhood acute lymphoblastic leukemia
54. A study of mitochondrial and nuclear transcription with cloned cDNA probes. Changes in the relative abundance of mitochondrial transcripts after stimulation of quiescent mouse fibroblasts
55. Multiplex PCR: Advantages, development, and applications
56. Quantitative RT-PCR to evaluate in vivo expression of multiple transgenes using a common intron
57. Reverse transcription-polymerase chain reaction-based methodology to quantify differential gene expression directly from microdissected regions of frozen tissue sections
58. Quantitative or semi-quantitative PCR: Reality versus myth
59. Real-time quantitative RT-PCR after laser-assisted cell picking
60. Regulation of hypoxanthine phosphoribosyltransferase, glyceraldehyde-3-phosphate dehydrogenase and beta-actin mRNA expression in porcine immune cells and tissues
61. Use of elevated reverse transcription reaction temperatures in RT-PCR
62. Quantitative RT-PCR: Pitfalls and potential
63. The hormonal regulation of the oestrogen receptor in rat liver: An interplay involving growth hormone, thyroid hormones and glucocorticoids
64. Direct random mutagenesis of gene-sized DNA fragments using polymerase chain reaction
65. Detection of colorectal carcinoma cells in circulating peripheral blood by reverse transcription-polymerase chain reaction targeting cytokeratin-20 mRNA
66. Cytokeratin 20 mRNA in peripheral venous blood of colorectal carcinoma patients
67. Expression of prostate-specific antigen and prostate-specific membrane antigen transcripts in blood cells: Implications for the detection of hematogenous prostate cells and standardization
68. Reverse transcriptase. The use of cloned Moloney murine leukemia virus reverse transcriptase to synthesize DNA from RNA
69. Improved quantitation of minimal residual disease in multiple myeloma using real-time polymerase chain reaction and plasmid-DNA complementarity determining region III standards
70. Polymerase chain reaction in the detection of micrometastases and circulating tumor cells
71. A novel method for real time quantitative RT-PCR
72. Molecular beacons: A new approach for semiautomated mutation analysis
73. Analysis of cytokine mRNA and DNA: Detection and quantitation by competitive polymerase chain reaction
74. Competitive PCR for quantitation of mRNA
75. Variation in expression of genes used for normalization of Northern blots after induction of cell proliferation
76. Reverse transcription, amplification and sequencing of poliovirus RNA by Taq DNA polymerase
77. Regulation of endothelial cell glyceraldehyde-3-phosphate dehydrogenase expression by hypoxia
78. Isothermal, in vitro amplification of nucleic acids by a multienzyme reaction modeled after retroviral replication
79. One-tube fluorogenic reverse transcription-polymerase chain reaction for the quantitation of feline coronaviruses
80. Comparative study of different standardization concepts in quantitative competitive reverse transcription-PCR assays
81. Complementary DNA for human T-cell cyclophilin
82. Modeling and analysis of competitive RT-PCR
83. Increased expression of glyceraldehyde-3-phosphate dehydrogenase in cultured astrocytes following exposure to manganese
84. Real time quantitative PCR
85. Detection of illegitimate transcripts of prostate-specific antigen mRNA in blood by reverse transcription-polymerase chain reaction
86. Increased analytical sensitivity of RT-PCR of PSA mRNA decreases diagnostic specificity of detection of prostatic cells in blood
87. Limitations imposed by heteroduplex formation on quantitative RT-PCR
88. Kinetic PCR analysis: Real-time monitoring of DNA amplification reactions
89. The polymerase chain reaction: Applications for the detection of foodborne pathogens
90. A simplified ribonuclease protection assay
91. Clinical application of reverse transcription-polymerase chain reaction for HIV infection
92. Improved results with PCR for chronic myeloid leukaemia
93. +-induced apoptosis but not necrosis of cultured cerebellar granule cells
94. Sensitive nested reverse transcription polymerase chain reaction detection of circulating prostatic tumor cells: Comparison of prostate-specific membrane antigen and prostate-specific antigen-based assays
95. Oxidative stress increases glyceraldehyde-3-phosphate dehydrogenase mRNA levels in isolated rabbit aorta
96. A simple and reliable protocol for the detection of apple stem grooving virus by RT-PCR and in a multiplex PCR assay
97. Amplification of human minisatellites by the polymerase chain reaction: Towards DNA fingerprinting of single cells
98. RNA quantitation by fluorescence-based solution assay: RiboGreen reagent characterization
99. Quality management and influential factors for the detection of single metastatic cancer cells by reverse transcriptase polymerase chain reaction
100. Differential quantitation of alternatively spliced messenger RNAs using isoform-specific real-time RT-PCR
101. Illegitimate transcription: Its use in the study of inherited disease
102. Quantification of mRNA by polymerase chain reaction (PCR) using an internal standard and a nonradioactive detection method
103. New prognostic factors in melanoma: mRNA tumour markers
104. Reliability of reverse transcription-polymerase chain reaction (RT-PCR)-based assays for the detection of circulating tumour cells: A quality-assurance initiative of the EORTC Melanoma Cooperative Group
105. Coupled reverse transcription-polymerase chain reaction (RT-PCR) technique is comparative, quantitative, and rapid: Uses in alcohol research involving low abundance mRNA species such as hypothalamic LHRH and GRF
106. Isolation of cloned Moloney murine leukemia virus reverse transcriptase lacking ribonuclease H activity
107. Highly sensitive and specific fluorescence reverse transcription-PCR assay for the pseudogene-free detection of beta-actin transcripts as quantitative reference
108. Quantification of cytokine mRNA expression by RT PCR in samples of previously frozen blood
109. Single blastomeres within human preimplantation embryos express different amounts of messenger ribonucleic acid for beta-actin and interleukin-1 receptor type I
110. Avoiding false positives with PCR
111. The detection of minimal numbers of contaminating epithelial tumor cells in blood or bone marrow: Use, limitations and future of RNA-based methods
112. Real-time fluorescence genotyping of factor V Leiden during rapid-cycle PCR
113. Allelic discrimination by nick-translation PCR with fluorogenic probes
114. Fluorescent detection techniques for real-time multiplex strand specific detection of Bacillus anthracis using rapid PCR
115. Seven-color, homogeneous detection of six PCR products
116. Quantitative comparison of mRNA levels in mammalian tissues: 28S ribosomal RNA level as an accurate internal control
117. Molecular beacon probes combined with amplification by NASBA enable homogeneous, real-time detection of RNA
118. Quantitative real-time PCR for the measurement of feline cytokine mRNA
119. Use of real-time PCR and molecular beacons to detect virus replication in human immunodeficiency virus type 1 infected individuals on prolonged effective antiretroviral therapy
120. Allelic discrimination using fluorogenic probes and the 5' nuclease assay
121. Oligonucleotides with fluorescent dyes at opposite ends provide a quenched probe system useful for detecting PCR product and nucleic acid hybridization
122. Mutation detection and single-molecule counting using isothermal rolling-circle amplification
123. Alternative splicing of pre-mRNA: Developmental consequences and mechanisms of regulation
124. Structure-specific endonucleolytic cleavage of nucleic acids by eubacterial DNA polymerases
125. Cell cycle regulation of the glyceraldehyde-3-phosphate dehydrogenase/uracil DNA glycosylase gene in normal human cells
126. Quantitative method to determine mRNA levels by reverse transcriptase-polymerase chain reaction from leukocyte subsets purified by fluorescence-activated cell sorting: Application to peripheral cannabinoid receptors
127. Multiplex detection of single-nucleotide variations using molecular beacons
128. High-throughput real-time reverse transcription-PCR quantitation of hepatitis C virus RNA
129. Development of a quantitative gene expression assay for Chlamydia trachomatis identified temporal expression of sigma factors
130. In situ visualization of messenger RNA for basic fibroblast growth factor in living cells
131. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter
132. Simultaneous screening for HBV DNA and HCV RNA genomes in blood donations using a novel TaqMan PCR assay
133. Semi-quantitative polymerase chain reaction analysis of cytokine and cytokine receptor gene expression during thymic ontogeny
134. Measuring infectious bursal disease virus RNA in blood by multiplex real-time quantitative RT-PCR
135. Rapid reverse transcription-PCR detection of hepatitis C virus RNA in serum by using the TaqMan fluorogenic detection system
136. Quantification of low-copy transcripts by continuous SYBR Green I monitoring during amplification
137. Pitfalls of processed pseudogenes in RT-PCR
138. Reverse transcription and DNA amplification by a Thermus thermophilus DNA polymerase
139. Molecular diagnostic detection of free cancer cells in the peritoneal cavity of patients with gastrointestinal and gynecologic malignancies
140. Rapid and reliable detection of N-ras mutations in acute lymphoblastic leukemia by melting curve analysis using LightCycler technology
141. An insulin response element in the glyceraldehyde-3-phosphate dehydrogenase gene binds a nuclear protein induced by insulin in cultured cells and by nutritional manipulations in vivo
142. Minilibraries constructed from cDNA generated by arbitrarily primed RT-PCR: An alternative to normalized libraries for the generation of ESTs from nanogram quantities of mRNA
143. Different real-time PCR formats compared for the quantitative detection of human cytomegalovirus DNA
144. Amplimers with 3'-terminal phosphorothioate linkages resist degradation by vent polymerase and reduce Taq polymerase mispriming
145. Detection of viable Listeria monocytogenes with a 5' nuclease PCR assay
146. Comparison of collagen gene expression in the developing chick embryo tendon and heart. Tissue and development time-dependent action of dexamethasone
147. The housekeeping gene glyceraldehyde-3-phosphate dehydrogenase is inappropriate as internal control in comparative studies between skin tissue and cultured skin fibroblasts using Northern blot analysis
148. Developments in quantitative PCR
149. Quantification of murine cytokine mRNAs using real time quantitative reverse transcriptase PCR
150. mRNA: Detection by in situ and northern hybridization
151. Survey and summary: Transcription by RNA polymerases I and III
152. Theoretical uncertainty of measurements using quantitative polymerase chain reaction
153. Thermostable DNA polymerases
154. Effect of inhibitors in clinical specimens on Taq and Tth DNA polymerase-based PCR amplification of influenza A virus
155. Variation of transferrin mRNA concentration in the rabbit mammary gland during the pregnancy-lactation-weaning cycle and in cultured mammary cells. A comparison with the other major milk protein mRNAs
156. Quantitative PCR: Theoretical considerations with practical implications
157. General principles of quantitative PCR
158. Design and testing of beta-actin primers for RT-PCR that do not co-amplify processed pseudogenes
159. Utilization of polymerase chain reaction technology in the detection of solid tumors
160. Multidrug resistance in human tumors - Molecular diagnosis and clinical significance
161. Wound macrophages express TGF-alpha and other growth factors in vivo: Analysis by mRNA phenotyping
162. Quantitative PCR
163. GAPDH acts as an inducible not constitutive gene in cultured endothelial cells
164. Alteration of glyceraldehyde-3-phosphate dehydrogenase activity and messenger RNA content by androgen in human prostate carcinoma cells
165. Product differentiation by analysis of DNA melting curves during the polymerase chain reaction
166. Quantification of alternatively spliced RUSH mRNA isoforms by QRT-PCR and IP-RPHPLC analysis: A new approach to measuring regulated splicing efficiency
167. Evidence of increased glyceraldehyde-3-phosphate dehydrogenase and fatty acid synthetase promoter activities in transiently transfected adipocytes from genetically obese rats
168. Optimization of the annealing temperature for DNA amplification in vitro
169. A faster ribonuclease protection assay
170. Polymerase chain reaction-based mRNA quantification using an internal standard: Analysis of oncogene expression
171. Increased sensitivity for the detection of malignant melanoma cells in peripheral blood using an improved protocol for reverse transcription-polymerase chain reaction
172. Spreadsheet software for thermodynamic melting point prediction of oligonucleotide hybridization with and without mismatches
173. Simultaneous TD/RT-PCR detection of cymbidium mosaic potexvirus and odontoglossum ringspot tobamovirus with a single pair of primers
174. Semi-automated fluorogenic PCR assays (TaqMan) for rapid detection of Escherichia coli O157:H7 and other Shiga toxigenic E. coli
175. TSH receptor status of thyroid neoplasms - TaqMan RT-PCR analysis of archival material
176. Use of avian myeloblastosis virus reverse transcriptase at high temperature for sequence analysis of highly structured RNA
177. Rapid identification of up to three Candida species in a single reaction tube by a 5' exonuclease assay using fluorescent DNA probes
178. PCR mimics: Competitive DNA fragments for use as internal standards in quantitative PCR
179. The detection of disseminated tumor cells in bone marrow from colorectal-cancer patients by a cytokeratin-20-specific nested reverse-transcriptase-polymerase-chain reaction is related to the stage of disease
180. Real time detection of DNA·RNA hybridization in living cells
181. Quantitative polymerase chain reaction does not improve preoperative prostate cancer staging: A clinicopathological molecular analysis of 121 patients
182. Quantitative RT-PCR: Limits and accuracy
183. Problems related to the interpretation of autoradiographic data on gene expression using common constitutive transcripts as controls
184. Analysis of gene expression in single oocytes and embryos by real-time rapid cycle fluorescence monitored RT-PCR
185. Molecular staging of prostate cancer: Dream or reality?
186. Housekeeping genes as internal standards: Use and limits
187. Rapid quantitation of gene therapy specific CFTR expression using the amplification refractory mutation system
188. Heterogeneity of primer extension products in asymmetric PCR is due both to cleavage by a structure-specific exo/endonuclease activity of DNA polymerases and to premature stops
189. Molecular beacons: Probes that fluoresce upon hybridization
190. Extremely sensitive, background-free gene detection using binary probes and beta replicase
191. Multicolor molecular beacons for allele discrimination
192. Construction of recombinant RNA templates for use as internal standards in quantitative RT-PCR
193. Multiplex detection of four pathogenic retroviruses using molecular beacons
194. Reproducibility of detection of tyrosinase and MART-1 transcripts in the peripheral blood of melanoma patients: A quality control study using real-time quantitative RT-PCR
195. Strand displacement amplification - An isothermal, in vitro DNA amplification technique
196. mRNA quantification by real time TaqMan polymerase chain reaction: Validation and comparison with RNase protection
197. Quantitation of mRNA by the polymerase chain reaction
198. A seminested RT-PCR assay for HER2/neu: Initial validation of a new method for the detection of disseminated breast cancer cells
199. Comparison of nested competitive RT-PCR and real-time RT-PCR for the detection and quantification of AML1/MTG8 fusion transcripts in t(8;21) positive acute myelogenous leukemia
200. Detection of rare mRNAs via quantitative RT-PCR
201. Development and validation of real-time quantitative reverse transcriptase-polymerase chain reaction for monitoring gene expression in cardiac myocytes in vitro
202. Accurate quantification of the mRNA of NMDAR1 splice variants measured by competitive RT-PCR
203. Rapid cycle DNA amplification: Time and temperature optimization
204. Automated polymerase chain reaction in capillary tubes with hot air
205. Minimizing the time required for DNA amplification by efficient heat transfer to small samples
206. Continuous fluorescence monitoring of rapid cycle DNA amplification
207. The LightCycler: A microvolume multisample fluorimeter with rapid temperature control
208. The effect of temperature and oligonucleotide primer length on the specificity and efficiency of amplification by the polymerase chain reaction
209. The UAS of the yeast GAPDH promoter consists of multiple general functional elements including RAP1 and GRF2 binding sites
210. Quantitative reverse transcription-PCR assay of the RNA component of human telomerase using the TaqMan fluorogenic detection system
211. Effects of fasting on the expression of gastrin, cholecystokinin, and somatostatin genes and of various housekeeping genes in the pancreas and upper digestive tract of rats
212. Quantitative RT-PCR for neuroendocrine studies. A minireview
213. Differential priming of RNA templates during cDNA synthesis markedly affects both accuracy and reproducibility of quantitative competitive reverse-transcriptase PCR
214. Two variants of quantitative reverse transcriptase PCR used to show differential expression of alpha-, beta- and gamma-fibrinogen genes in rat liver lobes
215. Direct comparison of GAPDH, beta-actin, cyclophilin, and 28S rRNA as internal standards for quantifying RNA levels under hypoxia
216. Limitations of reverse-transcriptase polymerase chain reaction analyses for detection of micrometastatic epithelial cancer cells in bone marrow