Methods in Enzymology Vol.306 Expression of Recombinant Genes in Eukaryotic Systems

Discussion in 'Methods in Enzymology Book Series' started by iubongda12345, Aug 3, 2016.

  1. iubongda12345

    iubongda12345 New Member

    Although microarrays show great promise as tools for genotyping, mapping, and resequencing, an equally important application for the microarray is measuring transcript abundance. Microarrays have been used to simultaneously measure the mRNA expression levels for every gene in Saccharomyces cerevisiae under several different growth conditions, to characterize the differences between normal and metastatic tissues and as a vehicle for gene discovery. This chapter describes the two basic types of microarrays, protocols for fluorescently labeling messenger RNA from eukaryotic cells for hybridization to microarrays, and considerations involved in experimental design and data analysis. Many variations on the protocols are feasible: poly(A) purification steps might be eliminated, different fluors may work as well or better, and polymerase chain reaction (PCR) amplification of cDNA may be possible. A potential problem for all microarray experiments is cross-hybridization. Because microarrays generally have longer probes than do oligonucleotide arrays, some have argued that their specificity is greater and that the potential for cross-hybridization is lower.
    • Series: Methods in Enzymology (Book 306)
    • Hardcover: 403 pages
    • Publisher: Academic Press; 1 edition (July 29, 1999)
    • Language: English
    • ISBN-10: 0121822079
    • ISBN-13: 978-0121822071
    • Product Dimensions: 9.3 x 6.4 x 1 inches
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    Last edited by a moderator: Jan 7, 2022

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