Enzymes of Molecular Biology (Methods in Molecular Biology by Michael M. Burrell PDF


By Michael M. Burrell

ISBN-10: 0896033228

ISBN-13: 9780896033221

Offers key details on a variety of enzymes well-known as instruments in molecular biology, supporting to lessen the time a scientist spends discovering the literature to get reactions to paintings successfully and permitting the nonenzymologist to layout an test. every one bankruptcy offers heritage info at the enzyme chosen and people parameters very important in its use, describes either the resource and alertness of the enzyme, and gives info at the measurement and constitution of the protein. particular parameters crucial for reaching an optimized response are mentioned, besides exemplary useful techniques an protocols.

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Extra info for Enzymes of Molecular Biology (Methods in Molecular Biology Vol 16)

Example text

DNA 12,5 15-5 19 86. , Reiss, A , and Adesmk, M. (1989) Construction of mutant and chimeric genes using the polymerase chain reactton. Nucleic Acids Res 17,723-733. Landgraf and Wolfes 87. Mole, S. , Iggo, R. , and Lane, D. P. (1989) Using the polymerase cham reaction to modify expression plasmtds for epitope mapping. Nuclerc Acids Res 17,33 19 88. Yon, J. and Fried, M (1989) Precise gene fusion by PCR. Nucleic Acids Res 17,4895. 89. Frye, R. A , Benz, C C , and Lm, E (1989) Detection of amplified oncogenes by differential polymerase chain reaction.

Hence, the product yield is dependent on the enzyme concentration (13) (Fig. 3). 4 02 n ” I I I I I 0 1 2 3 4 U taq polymerase Fig 3 Dependency of primer mcorporation on enzyme amounts a,22 cycles, n , 24 cycles, A, 26 cycles The initial template concentration hardly influences the product yield in the plateau phase. Even l-10 copies of target sequence will be amplified to give the plateau concentration after 40-50 cycles. g DNA. Therefore, the amplification has to reach the plateau phase, if the product is to be analyzed by gel electrophoresis and ethidium bromide staining.

Olsen, D B and Eckstein, F (1989) Incomplete primer extension during zn vitro DNA amplification catalyzed by Taq polymerase; explottatron for DNA sequencing Nucleic Acrds Res. 17,9613-9620. 85 Rochhtz, C F , Scott, G. , Dodson, J M , and Benz, C C. (1988) Use of the polymerase chain reaction technique to create base-specific ras oncogene mutations. DNA 12,5 15-5 19 86. , Reiss, A , and Adesmk, M. (1989) Construction of mutant and chimeric genes using the polymerase chain reactton. Nucleic Acids Res 17,723-733.

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Enzymes of Molecular Biology (Methods in Molecular Biology Vol 16) by Michael M. Burrell

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