By Michael M. Burrell
Offers key info on a variety of enzymes conventional as instruments in molecular biology, assisting to reduce the time a scientist spends gaining knowledge of the literature to get reactions to paintings successfully and permitting the nonenzymologist to layout an scan. each one bankruptcy supplies history details at the enzyme chosen and people parameters vital in its use, describes either the resource and alertness of the enzyme, and gives information at the dimension and constitution of the protein. particular parameters crucial for reaching an optimized response are mentioned, in addition to exemplary useful approaches an protocols.
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Extra resources for Enzymes of Molecular Biology (Methods in Molecular Biology Vol 16)
Sometimes it is difficult to detect a single mutation in longer fragments using denaturation gradient gel electrophoresis (DGGE). This drawback can be overcome by attachment of a 40-bp GC-rich stretch to the primers (60). The analysis of the PCR product, melting at a higher temperature, will enhance the sensitivity of this method. Mixed oligodeoxynucleotide primers designedon the basis of the amino acid sequenceof urate oxidase were used successfully to clone the gene for this enzyme (34). If the primers crosshybridize to repetitive sequences, even an optimal adaptation of the annealing temperature will not eliminate unspecific bands.
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