By P. D. Bridge, D. K. Arora, C. A. Reddy, R. P. Elander
Polymerase chain response (PCR), a robust procedure for amplifying DNA fragments, has had a tremendous effect on study all through biology. This e-book discusses the wide-ranging functions of PCR in natural and utilized mycology.
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Extra info for Applications of PCR in Mycology
Plant Molecular Biology 32, 947–957. , Rapley, R. R. (1992) Sequencing of PCR-amplified DNA. PCR Methods and Applications 1, 222–228. J. M. (1991) DNA amplification fingerprinting using very short arbitrary oligonucleotide primers. Biotechnology 9, 553–557. L. (1992) Rapid extraction of fungal DNA for PCR amplification. Nucleic Acids Research 20, 2380. S. G. (1993) Specificity, efficiency, and fidelity of PCR. PCR Methods and Applications 3, S18–S29. Chen, W. (1992) Restriction fragment length polymorphisms in enzymatically amplified ribosomal DNAs of three heterothallic Pythium species.
1994). 2 Source of DNA An extensive search of literature from the past 5 years shows that most instances of the use of PCR in the cloning of fungal genes have used genomic DNA as a template. Since neither high quantities nor high quality of DNA are needed, a number of simple and rapid protocols can be used for DNA extraction. , 1995). 3 Primer design Design of primers is one of the key steps in the successful cloning of a gene. However, no absolute rules can be given that will guarantee the amplification of the desired fragment in sufficient quantities.
1991; Hu, 1993; Biotechnology catalogue, Perkin Elmer, New Jersey, USA; Promega catalogue, Madison, Wisconsin, USA; Pfu DNA Polymerase Instruction Manual, Stratagene, La Jolla, California, USA, and references therein; data sheets for Vent DNA polymerase from NEB, New England, USA; Pwo DNA polymerase from Boehringer Mannheim, Germany). The table is therefore incomplete. c Expandase activity refers to the addition of a non-templated nucleotide to the 3′-end of blunt-ended DNA molecules. d Pfu DNA polymerase remains > 95% active following a 1 h incubation at 95°C.
Applications of PCR in Mycology by P. D. Bridge, D. K. Arora, C. A. Reddy, R. P. Elander