Literature DB >> 7606501

Site-directed mutagenesis by double polymerase chain reaction.

S Barik1.   

Abstract

A simple and versatile method of in vitro site-specific mutagenesis based on polymerase chain reaction (PCR) is described. The complete method required the use of three oligonucleotide primers and two PCRs. The product of the first PCR was used as one of the primers (megaprimer) in the second PCR. Essentially 100% of the final product incorporated the desired mutation. The various aspects of the procedure and its application is described in detail.

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Year:  1995        PMID： 7606501     DOI： 10.1007/BF02821329

Source DB:  PubMed          Journal:  Mol Biotechnol        ISSN： 1073-6085            Impact factor:   2.695


  7 in total

1.  Producing single-stranded DNA probes with the Taq DNA polymerase: a high yield protocol.

Authors:  U Finckh; P A Lingenfelter; D Myerson
Journal:  Biotechniques       Date:  1991-01       Impact factor: 1.993

2.  "Megaprimer" method of PCR: increased template concentration improves yield.

Authors:  S Barik; M S Galinski
Journal:  Biotechniques       Date:  1991-04       Impact factor: 1.993

3.  Rapid re-amplification of PCR products purified in low melting point agarose gels.

Authors:  C B Zintz; D C Beebe
Journal:  Biotechniques       Date:  1991-08       Impact factor: 1.993

4.  A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions.

Authors:  R Higuchi; B Krummel; R K Saiki
Journal:  Nucleic Acids Res       Date:  1988-08-11       Impact factor: 16.971

5.  Genomic amplification with transcript sequencing.

Authors:  E S Stoflet; D D Koeberl; G Sarkar; S S Sommer
Journal:  Science       Date:  1988-01-29       Impact factor: 47.728

6.  The "megaprimer" method of site-directed mutagenesis.

Authors:  G Sarkar; S S Sommer
Journal:  Biotechniques       Date:  1990-04       Impact factor: 1.993

7.  Novel non-templated nucleotide addition reactions catalyzed by procaryotic and eucaryotic DNA polymerases.

Authors:  J M Clark
Journal:  Nucleic Acids Res       Date:  1988-10-25       Impact factor: 16.971
  7 in total
  19 in total

1.  Altered pathway routing in a class of Salmonella enterica serovar Typhimurium mutants defective in aminoimidazole ribonucleotide synthetase.

Authors:  J L Zilles; T J Kappock; J Stubbe; D M Downs
Journal:  J Bacteriol       Date:  2001-04       Impact factor: 3.490

2.  Identification of specific residues in colicin E1 involved in immunity protein recognition.

Authors:  M Lindeberg; W A Cramer
Journal:  J Bacteriol       Date:  2001-03       Impact factor: 3.490

3.  Mutations in transhydrogenase change the fluorescence emission state of TRP72 from 1La to 1Lb.

Authors:  Karina Tveen Jensen; Giovanni Strambini; Margherita Gonnelli; Jaap Broos; J Baz Jackson
Journal:  Biophys J       Date:  2008-07-03       Impact factor: 4.033

4.  Mutational analysis of active-site residues in the Mycobacterium leprae RecA intein, a LAGLIDADG homing endonuclease: Asp(122) and Asp(193) are crucial to the double-stranded DNA cleavage activity whereas Asp(218) is not.

Authors:  Pawan Singh; Pankaj Tripathi; K Muniyappa
Journal:  Protein Sci       Date:  2010-01       Impact factor: 6.725

5.  Autoregulation of the MisR/MisS two-component signal transduction system in Neisseria meningitidis.

Authors:  Yih-Ling Tzeng; Xiaoliu Zhou; Shaojia Bao; Shuming Zhao; Corie Noble; David S Stephens
Journal:  J Bacteriol       Date:  2006-07       Impact factor: 3.490

6.  A rapid and efficient method for site-directed mutagenesis using one-step overlap extension PCR.

Authors:  A Urban; S Neukirchen; K E Jaeger
Journal:  Nucleic Acids Res       Date:  1997-06-01       Impact factor: 16.971

7.  Recruitment of the priming protein pTP and DNA binding occur by overlapping Oct-1 POU homeodomain surfaces.

Authors:  R N de Jong; M E Mysiak; L A T Meijer; M van der Linden; P C van der Vliet
Journal:  EMBO J       Date:  2002-02-15       Impact factor: 11.598

8.  A periplasmic location is essential for the role of the ApbE lipoprotein in thiamine synthesis in Salmonella typhimurium.

Authors:  B J Beck; D M Downs
Journal:  J Bacteriol       Date:  1999-12       Impact factor: 3.490

9.  Identification of essential lysines involved in substrate binding of vacuolar H+-pyrophosphatase.

Authors:  Chien-Hsien Lee; Yih-Jiuan Pan; Yun-Tzu Huang; Tseng-Huang Liu; Shen-Hsing Hsu; Ching-Hung Lee; Yen-Wei Chen; Shih-Ming Lin; Lin-Kun Huang; Rong-Long Pan
Journal:  J Biol Chem       Date:  2011-02-03       Impact factor: 5.157

10.  Mutations in the 3beta-hydroxysterol Delta24-reductase gene cause desmosterolosis, an autosomal recessive disorder of cholesterol biosynthesis.

Authors:  H R Waterham; J Koster; G J Romeijn; R C Hennekam; P Vreken; H C Andersson; D R FitzPatrick; R I Kelley; R J Wanders
Journal:  Am J Hum Genet       Date:  2001-08-22       Impact factor: 11.025
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