Improved Antibiotic-Resistance Cassettes Through Restriction Site Elimination Using Pfu DNA Polymerase PCR
نویسندگان
چکیده
منابع مشابه
Improved antibiotic-resistance cassettes through restriction site elimination using Pfu DNA polymerase PCR.
1.Eghtedarzadeh-Kondri, M., M.A. Walls and S.M. Glaser. 1997. Site-specific mutagenesis of immunoglobulin domains by multiple-fragment homologous recombination. BioTechniques 23:830-834. 2.Ho, S.N., H.D. Hunt, R.M. Horton, J.K. Pullen and L.R. Pease. 1989. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51-59. 3.Huff, J.P., B.J. Grant, C.A. Penning an...
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Here we describe a simple and cost-effective PCR-based method to introduce deletions, point mutations, frame-shift, or truncations with very high efficiency. The scope and additional applications of the method are discussed. Site-directed mutagenesis involves the introduction of mutations at the DNA level to alter the primary amino acid sequence of proteins (1,2). To approach the desired level ...
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A rapid method for efficiently generating site-directed mutations on a clean sequence background is described. This modification of the megaprimer PCR mutagenesis approach can be performed in one tube in less than 4.5 hours, and does not require purification of intermediate products. High fidelity of DNA sequence replication is obtained by employing Pfu DNA polymerase and limiting the total num...
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The replication fidelities of Pfu, Taq, Vent, Deep Vent and UlTma DNA polymerases were compared using a PCR-based forward mutation assay. Average error rates (mutation frequency/bp/duplication) increased as follows: Pfu (1.3 x 10(-6)) < Deep Vent (2.7 x 10(-6)) < Vent (2.8 x 10(-6)) < Taq (8.0 x 10(-6)) < < exo- Pfu and UlTma (approximately 5 x 10(-5)). Buffer optimization experiments indicated...
متن کاملRAC: Repository of Antibiotic resistance Cassettes
Antibiotic resistance in bacteria is often due to acquisition of resistance genes associated with different mobile genetic elements. In Gram-negative bacteria, many resistance genes are found as part of small mobile genetic elements called gene cassettes, generally found integrated into larger elements called integrons. Integrons carrying antibiotic resistance gene cassettes are often associate...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 1998
ISSN: 0736-6205,1940-9818
DOI: 10.2144/98255bm04