Book ChapterDOI
Transcription maps of adenovirus.
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This chapter discusses the techniques for the transcription mapping of adenovirus and two methods have been developed to map spliced Ad2 mRNAs that include the nuclease gel electrophoresis procedure and electron microscopy of RNA-DNA hybrids.Abstract:
Publisher Summary This chapter discusses the techniques for the transcription mapping of adenovirus. All true early mRNAs and proteins can be found in adenovirus 2 (Ad2) transformed cell lines. However, only a subset of these early gene products is required to establish a transformed state. With the transition from early to late phase of infection several new mRNAs appear in the cytoplasm that are transcribed from the same regions of the viral DNA as the early mRNAs, but which have novel sizes and sequence content. Two methods have been developed to map spliced Ad2 mRNAs that include (1) the nuclease gel electrophoresis procedure and (2) electron microscopy of RNA-DNA hybrids. The strengths and limitations of these two techniques are reviewed in the chapter. A summary transcription map of Ad2 is also presented.read more
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Real time quantitative PCR.
TL;DR: Unlike other quantitative PCR methods, real-time PCR does not require post-PCR sample handling, preventing potential PCR product carry-over contamination and resulting in much faster and higher throughput assays.
Journal ArticleDOI
A gene map of the human genome
Gregory D. Schuler,Mark S. Boguski,Elizabeth A. Stewart,Lincoln Stein,Gabor Gyapay,Kate Rice,Robert E. White,P. Rodriguez-Tomé,Amita Aggarwal,Eva Bajorek,S. Bentolila,B. B. Birren,Adam Butler,Andrew B. Castle,N. Chiannilkulchai,Angela M. Chu,C M Clee,Sid Cowles,P. J. R. Day,T. Dibling,N. Drouot,Ian Dunham,Simone Duprat,C. East,C A Edwards,Jun Fan,Nicole Y. Fang,Cécile Fizames,Christine Garrett,L. Green,David Hadley,Midori A. Harris,Paul Harrison,Shannon T. Brady,Andrew A. Hicks,E. Holloway,L. Hui,S. Hussain,C. Louis-Dit-Sully,J. Ma,A. MacGilvery,Christopher Mader,A. Maratukulam,Tara C. Matise,K. B. McKusick,Jean Morissette,Andrew J. Mungall,Delphine Muselet,H. C. Nusbaum,David C. Page,Ammon B. Peck,Shanti M. Perkins,Mark Piercy,Fawn Qin,John Quackenbush,S A Ranby,Tim Reif,Steve Rozen,C. Sanders,X. She,James Silva,Donna K. Slonim,Carol Soderlund,W.-L. Sun,P. Tabar,T. Thangarajah,Nathalie Vega-Czarny,Douglas Vollrath,S. Voyticky,T. E. Wilmer,Xiao-Yu Wu,Mark Raymond Adams,Charles Auffray,Nicole A.R. Walter,Rhonda Brandon,Anindya Dehejia,Peter N. Goodfellow,R. Houlgatte,James R. Hudson,Susan E. Ide,K. R. Iorio,Wha‐Young Lee,N. Seki,Takahiro Nagase,K. Ishikawa,N. Nomura,Cheryl Phillips,Mihael H. Polymeropoulos,Mina Sandusky,Karin Schmitt,Richard Berry,K. Swanson,R. Torres,J. C. Venter,James M. Sikela,Jacques S. Beckmann,Jean Weissenbach,Richard M. Myers,David R. Cox,Michael R. James,David Bentley,Panos Deloukas,Eric S. Lander,Thomas J. Hudson,Thomas J. Hudson +104 more
TL;DR: The gene map unifies the existing genetic and physical maps with the nucleotide and protein sequence databases in a fashion that should speed the discovery of genes underlying inherited human disease.
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Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae.
Mark Johnston,Ronald W. Davis +1 more
TL;DR: The GAL1 and GAL10 genes of Saccharomyces cerevisiae are divergently transcribed, with 606 base pairs of DNA separating their transcription initiation sites as mentioned in this paper.
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Cloning and analysis of the promoter region of the erythromycin resistance gene (ermE) of Streptomyces erythraeus.
TL;DR: It is suggested that these sites represent a class of vegetatively expressed Streptomyces promoter that is utilised by a form of RNA polymerase holoenzyme that also recognizes typical promoters of other bacterial genera.
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Quantitation of Dihydropyrimidine Dehydrogenase Expression by Real-Time Reverse Transcription Polymerase Chain Reaction
TL;DR: This method provides the first high-throughput, reproducible, and sensitive technique capable of determining DPD mRNA expression levels in nanogram amounts of total RNA, and is validated by quantitative comparison to Northern blot analysis in five tissues.
References
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Journal ArticleDOI
Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I
TL;DR: Labeled DNAs (and restriction endonuclease fragments derived from them) are useful probes for detecting rare homologous sequences by in situ hybridization and reassociation kinetic analysis.
Journal ArticleDOI
Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids
Arnold J. Berk,Phillip A. Sharp +1 more
TL;DR: A simple and sensitive method for detecting, sizing and mapping RNA transcripts from viral or cloned DNAs has been developed and used to examine the cytoplasmic transcripts produced during the early phase of adenovirus 2 (Ad2) infection of HeLa cells.
Journal ArticleDOI
Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis
Journal ArticleDOI
Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels.
TL;DR: Electrophoresis in alkaline gels can provide accurate molecular weights for linear, single-Stranded DNAs, and should be useful in analyzing DNA for single-strand breaks, depurinations or topological differences such as ring forms.
Journal ArticleDOI
Kinetics of renaturation of DNA.
James G. Wetmur,Norman Davidson +1 more
TL;DR: It is proposed that the mechanism of the reaction involves the joining of short, homologous sites on the two strands followed by a fast, reversible zippering reaction with forward rate constant kt, which explains the temperature and the GC dependence.