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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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Duplicate polyphenol oxidase genes on barley chromosome 2H and their functional differentiation in the phenol reaction of spikes and grains

TL;DR: Observations demonstrate that PPO1 is the major determinant controlling the phenol reaction of awns, and Comparisons of P PO1 single mutants and the PPO 1PPO2 double mutant indicate that P PO2 controls the Phenol reaction in the crease on the ventral side of caryopses.
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Characterisation of Muta™Mouse λgt10-lacZ transgene: evidence for in vivo rearrangements.

TL;DR: The model for λgt10-lacZ transgene organisation, and the PCR-based methods for assessing copy number, integrity and rearrangements, potentially extends the use of Muta™Mouse construct for direct, genomic-type assays that detect the effects of clastogens and aneugens, without depending on an E.coli host, for reporting effects.
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Biochemical genetics of glucosinolate modification in Arabidopsis and Brassica

TL;DR: Fine mapping of the glucosinolate biosynthesis gene OHP in an Arabidopsis thaliana Columbia × Landsberg erecta RI line population positioned the gene within 54 kb of DNA on chromosome IV, identifying a family of genes encoding 2-oxoglutarate-dependent dioxygenases in this region.
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Identification and function of a polyketide synthase gene responsible for 1,8-dihydroxynaphthalene-melanin pigment biosynthesis in Ascochyta rabiei

TL;DR: It is demonstrated that A. rabiei uses a DHN-melanin pathway for pigmentation of pycnidia and this molecule may protect pycnidiospores from UV irradiation, and an orthologous PKS1 gene from Bipolaris oryzae into the genome of the mutants complemented the dysfunctional ArP KS1 gene.
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Development of a real-time PCR assay targeting the sporulation gene, spo0A, for the enumeration of thermophilic bacilli in milk powder.

TL;DR: With this method the detection of thermophilic bacilli in milk powder can be accomplished within 1 h and the detection limit for reconstituted and inoculated milk was 80 vegetative cfu ml (-1) and 640 spores ml(-1), respectively.
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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