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M. Dagert

Bio: M. Dagert is an academic researcher. The author has contributed to research in topics: Incubation & Recombinant DNA. The author has an hindex of 1, co-authored 1 publications receiving 1347 citations.

Papers
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Journal ArticleDOI
01 Jan 1979-Gene
TL;DR: Escherichia coli cells are 4--6 times more transformable and 20--30 times more competent after 24 h incubation in cold calcium chloride than immediately after calcium chloride treatment.

1,354 citations


Cited by
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Journal ArticleDOI
TL;DR: Competition with both transforming and non-transforming plasmids indicates that each cell is capable of taking up many DNA molecules, and that the establishment of a transformation event is neither helped nor hindered significantly by the presence of multiple plasmid molecules.

11,144 citations

Journal ArticleDOI
01 Jan 1990-Gene
TL;DR: The conditions for preparing competent Escherichia coli cells are re-evaluated and a simple and efficient method (SEM) for plasmid transfection is established and these competent cells are particularly useful for construction of high-complexity cDNA libraries with a minimum expenditure of mRNA.

2,254 citations

Journal ArticleDOI
TL;DR: This sequence allowed the determination of the hitherto unknown primary structure of rat GAPDH which is 333 aminoacids long and revealed a high degree of sequence conservation at both nucleotide and protein levels.
Abstract: We have isolated and sequenced a full-length cDNA clone encoding rat glyceraldehyde-3-phosphate-dehydrogenase (GAPDH, E.C.1.2.1.12). The entire mRNA is 1269 nucleotides long exclusive of poly(A) and contains respectively 71 and 196 bases of 5' and 3' non-coding regions. Primer extension as well as S1 nuclease protection experiments clearly established that a single (or at least a highly prominent) GAPDH mRNA species is expressed in all rat tissues examined. This sequence allowed the determination of the hitherto unknown primary structure of rat GAPDH which is 333 aminoacids long. Comparison between GAPDH sequences from rat, man and chicken revealed a high degree of sequence conservation at both nucleotide and protein levels.

2,029 citations

Book ChapterDOI
TL;DR: Oligonucleotide-directed in vitro mutagenesis provides a means to alter a defined site within a region of cloned DNA, from the definition of functional DNA sequences to the construction of new proteins.
Abstract: Publisher Summary Oligonucleotide-directed in vitro mutagenesis provides a means to alter a defined site within a region of cloned DNA. This powerful technique has many far-reaching applications, from the definition of functional DNA sequences to the construction of new proteins. The method stemmed from the combination of a number of recent discoveries and observations about nucleic acids but the basic principle involves the enzymic extension by E. coli DNA polymerase of an oligonucleotide primer hybridized to a single-stranded circular template. More recently, phage M13 and M13 derived vectors have been used in a number of oligonucleotide mutagenesis experiments. Vectors derived from single-stranded phage, such as M13 and fd, are more convenient for oligonucleotide-directed mutagenesis than double-stranded vectors, because isolation of pure single-stranded circular template DNA in these systems is quite simple. This chapter describes a mutagenesis procedure that is simple, versatile, and efficient. The chapter also explains the targets and vehicles utilized in a number of recent mutagenesis experiments.

1,240 citations

Journal ArticleDOI
01 Jul 1983-Cell
TL;DR: The screened cDNA library for gene sequences that are regulated by platelet-derived growth factor (PDGF) in BALB/c-3T3 cells indicates that these sequences correspond to "early genes" which are not induced as a consequence of cell growth, but rather are directly regulated by PDGF.

863 citations