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Expression Genetics: Accelerated and High-Throughput Methods

  •  ISBN-10 1-881299-24-4

  •  ISBN-13 978-1-881299-24-0



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publication info   M. McClelland and A. Pardee, ed., Eaton Publishing, 1999, 385 pp., soft cover
mfr. no.   Eaton Publishing


General description

The methods included here, i.e., mammalian expression vectors, would come into play once regulation of a gene has been discovered by differential display. Advances in alternatives to differential display are also presented, including AFLP, arrays, protein display, and the two-hybrid system. These 37 papers selected from the journal BioTechniques along with their accompanying updates emphasize technologies that accelerate screening and selection assays.

Table of Contents

Section I. Making Arrays
1. Inexpensive handheld device for construction of high-density nucleic acid arrays,
2. Adapting the Biomek 2000 Laboratory Automation Work-station for printing DNA microarrays,
3. Parallel production of oligonucleotide arrays using membranes and reagent jet printing,
4. Agarose-embedded tissue arrays for histologic and genetic analysis,
Section II. High-Throughput DNA Screening
5. Using oligonucleotide probe arrays to access genetic diversity,
6. Direct hybridization of large-insert genomic clones on high-density gridded cDNA filter arrays, .
7. TurboPrep II: an inexpensive, high-throughput plasmid template preparation protocol,
Section III. High-Throughput RNA Screening: A. RNA Preparation
8. Amplification of mRNAs from single, fixed, TUNEL-positive cells,
9. Laser capture microdissection of single cells from complex tissues,
B. Fingerprinting Methods:
10. Non-radioisotopic AFLP method using PCR primers fluorescently labeled with Cy-5,
11. Modification of the AFLP protocol applied to honeybee (Apis mellifera L.) DNA,
12. Chemiluminescent detection of AFLP markers,
13. DAF optimization using Taguchi methods and the effect of thermal cycling parameters on DNA amplification,
C. Subtraction:
14. High-stringency subtraction for identification of differentially regulated cDNA clones,
15. Differential screening of a subtracted cDNA library: a method to search for genes preferentially expressed in multiple tissues,
16. Isolation of differentially expressed genes by combining representational difference analysis (RDA) and cDNA library arrays,
D. cDNA Array Applications:
17. High-throughput cDNA screening utilizing a low order neural network filter,
18. Nonradioactive detection of differentially expressed genes using complex RNA or DNA hybridization probes, .
19. Representative cDNA libraries and their utility in gene expression profiling,
E. Quantitative RT-PCR:
20. High-throughput RT-PCR analysis of multiple transcripts using a micro-plate RNA isolation procedure,
21. One-step fluorescent probe product-enhanced reverse transcriptase assay,
Section IV. High-throughput Protein Screening A. Display of Proteins:
22. Flow cytometric quantification of surface-displayed recombinant receptors on Staphylococci,
23. Gene VIII-based, phage-display vectors for selection against complex mixtures of ligands, .
24. Biopanning phage display libraries using magnetic beads vs. polystyrene plates,
25. Fusion proteins could generate false positives in peptide phage display,
26. Phosphorylation-directed antibodies in high-flux screens for compounds ......, .
B. Two-Hybrid Selection
27. Mammalian two-hybrid system: a complementary approach to the yeast two-hybrid system,
28. Development of a yeast trihybrid screen using stable yeast strains and regulated protein expression,
Section V. Functional Analysis: Expression Vectors A. Reporters
29. Use of a dicistronic expression cassette encoding the green fluorescent protein .......
30. Dual-function reporter protein for analysis of gene expression in living cells,
31. Fusion of green fluorescent protein with the Zeocin-resistance marker ...
32. Green fluorescent protein tag for studies of drug-induced translocation ....
33. Fission yeast expression vectors adapted for positive identification.....
B. Retroviral Vectors
34. Tracking and quantitation of retroviral-mediated transfer using ......
35. n geo, combined selection and reporter gene ......
36. Retroviral gene transfer in chrondrogenic limb bud micromass cultures,
37. High-efficiency gene transfer .......

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