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Open AccessJournal ArticleDOI

Genetic Applications of an Inverse Polymerase Chain Reaction

Howard Ochman, +2 more
- 01 Nov 1988 - 
- Vol. 120, Iss: 3, pp 621-623
TLDR
The feasibility of IPCR is shown by amplifying the sequences that flank an IS1 element in the genome of a natural isolate of Escherichia coli.
Abstract
A method is presented for the rapid in vitro amplification of DNA sequences that flank a region of known sequence. The method uses the polymerase chain reaction (PCR), but it has the primers oriented in the reverse direction of the usual orientation. The template for the reverse primers is a restriction fragment that has been ligated upon itself to form a circle. This procedure of inverse PCR (IPCR) has many applications in molecular genetics, for example, the amplification and identification of sequences flanking transposable elements. In this paper we show the feasibility of IPCR by amplifying the sequences that flank an IS1 element in the genome of a natural isolate of Escherichia coli.

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Journal ArticleDOI

Overexpression of Penicillin V Acylase from Streptomyces lavendulae and Elucidation of Its Catalytic Residues

TL;DR: The pva gene from Streptomyces lavendulae, encoding a novel penicillin V acylase (SlPVA), has been isolated and characterized and is classified as a new member of the Ntn-hydrolase superfamily, which belongs to a novel subfamily of acylases that recognize substrates with long hydrophobic acyl chains and have biotechnological applications in semisynthetic antifungal production.
Patent

Nucleic acid based data storage

TL;DR: In this paper, compositions, devices, systems and methods for the generation and use of biomolecule-based information for storage are described, including methods and systems for efficient transfer of preselected polynucleotides from a storage structure for reading stored information.
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Genome wide functional genetics in haploid cells

TL;DR: Haploid genetics has thus become a powerful alternative to RNAi or CRISPR based screens and open the possibility of combining the power of a haploid genome with pluripotency of embryonic stem cells to uncover fundamental biological processes in defined cell types at a genomic scale.
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The third type III module of human fibronectin mediates cell adhesion and migration.

TL;DR: This study demonstrated that the third FN type III repeat and several fragments containing the repeat promote cell spreading and migration of human dermal fibroblasts (HDFs), whereas the fourth repeat (FnIII4) did not.
Journal ArticleDOI

Phenotypic characterization of transposon-inserted mutants of Clostridium proteoclasticum B316T using extracellular metabolomics.

TL;DR: In this paper, metabolic footprint analysis was applied to characterize Tn 916 -inserted mutants of a hemicellulose-degrading rumen bacterium grown on complex culture medium.
References
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Journal ArticleDOI

Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase

TL;DR: A thermostable DNA polymerase was used in an in vitro DNA amplification procedure, the polymerase chain reaction, which significantly improves the specificity, yield, sensitivity, and length of products that can be amplified.
Journal ArticleDOI

Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.

TL;DR: Two new methods were used to establish a rapid and highly sensitive prenatal diagnostic test for sickle cell anemia, using primer-mediated enzymatic amplification of specific beta-globin target sequences in genomic DNA, resulting in the exponential increase of target DNA copies.
Book ChapterDOI

Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction.

TL;DR: A method whereby a nucleic acid sequence can be exponentially amplified in vitro is described in the chapter, and the possibility of utilizing a heat-stable DNA polymerase is explored so as to avoid the need for addition of new enzyme after each cycle of thermal denaturation.
Journal ArticleDOI

Direct cloning and sequence analysis of enzymatically amplified genomic sequences

TL;DR: A method is described for directly cloning enzymatically amplified segments of genomic DNA into an M13 vector for sequence analysis and promises to be a rapid method for obtaining reliable genomic sequences from nanogram amounts of DNA.
Journal ArticleDOI

DNA typing from single hairs

TL;DR: Three different means of DNA typing are used for the determination of amplified DNA fragment length differences, hybridization with allele-specific oligonucleotide probes, and direct DNA sequencing on single human hairs to detect genetically variable mitochondrial and nuclear DNA sequences.
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