Home Nonfiction 7 • A Beginner’s Guide to Microarrays - download pdf or read online

A Beginner’s Guide to Microarrays - download pdf or read online

By Kathryn Aboytes, Jason Humphreys, Sonya Reis (auth.), Eric M. Blalock (eds.)

ISBN-10: 1441987606

ISBN-13: 9781441987600

ISBN-10: 1461346843

ISBN-13: 9781461346845

A Beginner's consultant to Microarrays addresses audiences - the center facility supervisor who produces, hybridizes, and scans arrays, and the elemental study scientist who should be acting the research and examining the consequences. consumer pleasant insurance and particular protocols are supplied for the technical steps and methods considering many elements of microarray expertise, together with:

-Cleaning and coating glass slides,
-Designing oligonucleotide probes,
-Constructing arrays for the detection and quantification of other bacterial species,
-Preparing recognizing ideas,
-Troubleshooting recognizing difficulties,
-Setting up and operating a middle facility,
-Normalizing history sign and controlling for systematic variance,
-Designing experiments for max influence,
-Analyzing information with statistical approaches,
-Clustering info with machine-learning protocols.

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Extra info for A Beginner’s Guide to Microarrays

Sample text

2002). Thiol Chemistry Thiols, like amines, are nucleophilic agents that react with electrophiles. Unlike amines however, thiols undergo facile redox reactions that have been used extensively for b ioconjugations. Thiol-modified oligonucleotides can be immobilized onto a glass support by nucleophilic attack, heterobifunctional crosslinkers or thiolldisulfide exchange reactions. , 2000). g. acrylic moieties) to form Michael addition product thioethers (Figure 15). Rehman et aI. (1999) and Lindroos et aI.

This method allows direct coupling of stable 5' disulfide-modified oligonucleotides onto mercaptosilaneactivated glass surfaces without pretreatment. , 2000). The silanized ologonucleotides were prepared as 5' thioated DNA-thiosilane mixed disulfides. , 2000). Stripping and re-probing was also demonstrated. Lindroos et aI. (2001) compared silanized nucleic acids to a variety of other covalent attachment strategies including isothiocyanate, aldehyde, acrydite and thiol chemistries. , 1999). Rehman and coworkers (1999), in their desire to develop a stable linkage for solid phase peR, took advantage of this to develop a covalent-thermostable oligonucleotide attachment method.

Phosphate: R = DNA, R' = amine modified surface. B. EDC structure. EDe The use ofEDC is an older method, initially described by Gilham (1968) to attach DNA to cellulose membranes via the 5' -phosphate group. It is used to covalently attach nucleic acids to solid supports, and involves the activation of a an acid moiety (carboxylic or phosphoric X= C, P(OH) Figure 13A respectively) with the water soluble diimide l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride, EDC (Figure 13B). Subsequent reaction with an amine forms stable amide bonds (Ghosh and Musso, 1987; Hermanson, 1996).

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A Beginner’s Guide to Microarrays by Kathryn Aboytes, Jason Humphreys, Sonya Reis (auth.), Eric M. Blalock (eds.)

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