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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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Journal ArticleDOI

Transposition of Tnr1 in rice genomes to 5'-PuTAPy-3' sites, duplicating the TA sequence.

TL;DR: Data is presented that may suggest that Tnr1 is a defective form of an autonomous transposable element of rice species with genome types other than AA.
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Two Transsulfurylation Pathways in Klebsiella pneumoniae

TL;DR: It is reported that the related enteric bacterium Klebsiella pneumoniae encodes genes for both transsulfurylation pathways; genetic and biochemical analyses show that they are coordinately regulated to prevent futile cycling.
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Comparative Structural and Molecular Characterization of Ribitol-5-Phosphate-Containing Streptococcus oralis Coaggregation Receptor Polysaccharides

TL;DR: Findings identify genetic markers for the different ribitol-5-phosphate-containing types of RPS present in S. oralis and establish a close relationship between these polysaccharides and serogroup 10 CPSs of S. pneumoniae.
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The enzymes with benzil reductase activity conserved from bacteria to mammals.

TL;DR: It is demonstrated that many B. cereus strains had benzil reductase activity in vivo, and that the benzil reducectases shared 94-100% amino acid identities, which will be utilized to produce important chiral compounds.
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Resurrection of a Global, Metagenomically Defined Gokushovirus

TL;DR: It is demonstrated that virions can reliably infect and lysogenize hosts by hijacking a conserved chromosome-dimer resolution system, and that even partial motifs enable phages to persist cytoplasmically without leading to collapse of their host culture.
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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