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Literature DB >> 2517212

Rapid one-step characterization of recombinant vectors by direct analysis of transformed Escherichia coli colonies.

Abstract

We have developed a polymerase chain reaction (PCR) based procedure for rapidly analyzing recombinant vectors in whole bacterial cells. No purification, restriction mapping or sequencing of vectors is required and the results are available within 6 hours. Whole cells are added to a PCR mix that is designed to amplify DNA only if the correct insert is present in the required orientation. The presence of an appropriately sized band on an agarose gel is indicative of a correct clone.

Entities: Chemical Species

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Year: 1989 PMID： 2517212

Source DB: PubMed Journal: Biotechniques ISSN： 0736-6205 Impact factor: 1.993

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Cited

19 in total

1. Measurement of locus copy number by hybridisation with amplifiable probes.

Authors: J A Armour; C Sismani; P C Patsalis; G Cross
Journal: Nucleic Acids Res Date: 2000-01-15 Impact factor: 16.971

2. Reactivation of insertionally inactivated Shiga toxin 2 genes of Escherichia coli O157:H7 caused by nonreplicative transposition of the insertion sequence.

Authors: M Kusumoto; Y Nishiya; Y Kawamura
Journal: Appl Environ Microbiol Date: 2000-03 Impact factor: 4.792

8. Isolation and characterization of a second exe operon required for extracellular protein secretion in Aeromonas hydrophila.

Authors: R Jahagirdar; S P Howard
Journal: J Bacteriol Date: 1994-11 Impact factor: 3.490

9. Mapping of neutralizing epitopes on Renibacterium salmoninarum p57 by use of transposon mutagenesis and synthetic peptides.

Authors: Gregory D Wiens; Jennifer Owen
Journal: Appl Environ Microbiol Date: 2005-06 Impact factor: 4.792

10. Isolation and analysis of eight exe genes and their involvement in extracellular protein secretion and outer membrane assembly in Aeromonas hydrophila.

Authors: S P Howard; J Critch; A Bedi
Journal: J Bacteriol Date: 1993-10 Impact factor: 3.490

Rapid one-step characterization of recombinant vectors by direct analysis of transformed Escherichia coli colonies.

1. Measurement of locus copy number by hybridisation with amplifiable probes.

2. Reactivation of insertionally inactivated Shiga toxin 2 genes of Escherichia coli O157:H7 caused by nonreplicative transposition of the insertion sequence.

3. Allosteric activation of kinases: design and application of RapR kinases.

4. Rapid method for processing soil samples for polymerase chain reaction amplification of specific gene sequences.

5. Direct sequencing of unpurified PCR-amplified DNA by semi-exponential cycle sequencing (SECS).

6. Origin and evolution of the slime molds (Mycetozoa)

7. Magnetic bead capture of cDNAs from double-stranded plasmid cDNA libraries.

8. Isolation and characterization of a second exe operon required for extracellular protein secretion in Aeromonas hydrophila.

9. Mapping of neutralizing epitopes on Renibacterium salmoninarum p57 by use of transposon mutagenesis and synthetic peptides.

10. Isolation and analysis of eight exe genes and their involvement in extracellular protein secretion and outer membrane assembly in Aeromonas hydrophila.