Genetic Applications of an Inverse Polymerase Chain Reaction
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.read more
Citations
More filters
Journal ArticleDOI
Production of transgenic lily plants by Agrobacterium-mediated transformation.
TL;DR: A system for the production of transgenic plants was developed for the Oriental hybrid lily by Agrobacterium-mediated genetic transformation and verified to be transgenic by GUS histochemical assay and inverse PCR analysis.
Journal ArticleDOI
Ac transposition from a T-DNA can generate linked and unlinked clusters of insertions in the tomato genome.
Brian I. Osborne,Catherine Corr,James P. Prince,Reinhard Hehl,Steven D. Tanksley,Sheila McCormick,Barbara Baker +6 more
TL;DR: The distribution of transposed Acs around the T-DNA and at locations unlinked to theT-DNA indicates that Ac transposes to linked and unlinked sites in tomato as it does in maize.
Journal ArticleDOI
Isolation and sequence of omcA, a gene encoding a decaheme outer membrane cytochrome c of Shewanella putrefaciens MR-1, and detection of omcA homologs in other strains of S. putrefaciens.
Judith M. Myers,Charles R. Myers +1 more
TL;DR: The sequence of the omcA gene, which encodes a decaheme cytochrome c that is localized to the outer membrane (OM) of Shewanella putrefaciens MR-1, was determined and suggested the presence of a hydrophobic leader sequence which is cleaved during translocation of the protein to the OM.
Journal ArticleDOI
Capture PCR: efficient amplification of DNA fragments adjacent to a known sequence in human and YAC DNA.
TL;DR: In this article, a linker is added to the ends by ligation and multiple extension reactions are performed using a biotinylated primer derived from the known sequence, permitting the subsequent isolation of extension products on a streptavidin-coated support.
Journal ArticleDOI
Characterization of bacteriophage P1 library containing inserts of Drosophila DNA of 75-100 kilobase pairs.
TL;DR: The P1 cloning system has general utility in molecular genetics and may provide an important intermediate level of resolution in physical mapping of the Drosophila genome.
References
More filters
Journal ArticleDOI
Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase
Randall Keichi Saiki,David H. Gelfand,Susanne Stoffel,Stephen J. Scharf,Russell Higuchi,Glenn Thomas Horn,Kary B. Mullis,Henry A. Erlich +7 more
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.
Randall Keichi Saiki,Stephen J. Scharf,Fred A. Faloona,Kary B. Mullis,Glenn Thomas Horn,Henry A. Erlich,Norman Arnheim +6 more
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.
Kary B. Mullis,Fred A. Faloona +1 more
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.