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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

Structural basis for the methylation of A1408 in 16S rRNA by a panaminoglycoside resistance methyltransferase NpmA from a clinical isolate and analysis of the NpmA interactions with the 30S ribosomal subunit

TL;DR: Experimental and theoretical analyses suggest that NpmA flips out the target nucleotide A1408 to carry out the methylation of kanamycin, which poses a threat to the successful use of aminoglycosides in clinical practice.
Journal ArticleDOI

Simultaneous amplification of 5′ and 3′ cDNA ends based on template-switching effect and inverse PCR

TL;DR: A new approach for simultaneously amplifying the 5' and 3' ends of a desired cDNA is described, which is reliable and easy to perform, and very useful for isolating cDNA species of rare transcripts.
Journal ArticleDOI

Albino Leaf 2 is involved in the splicing of chloroplast group I and II introns in rice

TL;DR: The Albino Leaf 2 (AL2) gene is involved in the splicing of chloroplast group I and II introns and is responsible forchloroplast development in rice.
Patent

Methods of Identification Using Methylation of CPG

TL;DR: In this article, the materials and methods for the identification of methylated nucleotides in samples of genomic DNA were described, and the present invention also relates to methods of diagnosis of specific conditions by identification of specific methylated DNA.
Journal ArticleDOI

Inverse polymerase chain reaction for cloning cellular sequences adjacent to integrated hepatitis B virus DNA in hepatocellular carcinomas

TL;DR: By employing the inverse polymerase chain reaction (IPCR) method, two adjacent cellular sequences were cloned from 11 HBV integrants in three HCCs and one of the flanking cellular sequences was identified as the human 28S rRNA gene, the other was not found homologous to any known human sequences.
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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