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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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Citations
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Characterization of Brachyury genes in the dogfish S. canicula and the lamprey L. fluviatilis. Insights into gastrulation in a chondrichthyan.

TL;DR: The results suggest that gastrulation in the dogfish and in some amniotes shares striking similarities despite the phylogenetic distance between these species, which supports the hypothesis that the extensively divergent morphologies of gastrulae among vertebrates largely result from adaptations to the presence of yolk.
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Molecular characterization of the glutathione-dependent formaldehyde dehydrogenase gene FLD1 from the methylotrophic yeast Pichia methanolica

TL;DR: Molecular characterization of the FLD1 gene, which encodes glutathione‐dependent formaldehyde dehydrogenase (FLD), from the methylotrophic yeast Pichia methanolica, suggests thatFLD1 is involved in the detoxification of formaldehyde in meethanol metabolism.
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Comparative structural and functional analysis of two octaheme nitrite reductases from closely related Thioalkalivibrio species

TL;DR: A comparative structural and functional analysis of two homologous octaheme nitrite reductases from closely related Thioalkalivibrio species was performed and it was shown that both enzymes have similar catalytic properties, owing to high structural similarity.
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PRISE (PRImer SElector): software for designing sequence-selective PCR primers.

TL;DR: PRImer Selector is a new software package that implements several features that improve and streamline the design of sequence-selective PCR primers, and can be used to design primers with a range of specificities, targeting individual sequences as well as diverse assemblages of genes.
Journal ArticleDOI

Genomic structure and differential expression of two tandem-arranged GSTZ genes in rice

TL;DR: Using rice culture cells, the respective responses of OsGSTZ1 and OsG STZ2 genes against various treatments by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) suggested that the expression of Os gSTZ 1 and Os GSTZ 2 genes are differentially regulated in the culture cell of rice.
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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