Get Fluorescence In Situ Hybridization (FISH) - Application PDF

By Thomas Liehr

ISBN-10: 3540705805

ISBN-13: 9783540705802

This FISH program consultant presents an outline of the foundations and the elemental recommendations of fluorescence in situ hybridization (FISH) and primed in situ hybridization (PRINS), that are effectively used to check many features of genomic habit and adjustments. In 36 chapters, contributed by way of foreign specialists of their specific box, the these days a number of methods and purposes of the robust options are offered and exact protocols are given. defined listed below are tools utilizing numerous phone forms and tissues in addition to various organisms, similar to mammalians, bugs, vegetation and microorganisms. Multicolor FISH systems and specified functions corresponding to the characterization of marker chromosomes, breakpoints, cryptic aberrations, nuclear structure and epigenetic adjustments, in addition to (array-based) comparative genomic hybridization reviews are provided. total, the means of selection is brought for unmarried mobile research in human genetics, microbiology, animal and plant sciences.

Show description

Read Online or Download Fluorescence In Situ Hybridization (FISH) - Application Guide PDF

Best cell biology books

Current Protocols Essential Laboratory Techniques - download pdf or read online

The newest identify from the acclaimed present Protocols sequence, present Protocols crucial Laboratory options, 2e offers the hot researcher with the talents and realizing of the elemental laboratory methods essential to run winning experiments, resolve difficulties, and develop into a efficient member of the fashionable existence technology laboratory.

Get Melanoma Development: Molecular Biology, Genetics and PDF

The ebook offers accomplished insights into molecular adjustments in malignant cancer. the final mechanisms of cancer development and improvement are defined, in addition to new learn findings. Our present wisdom at the molecules excited about mobilephone transformation and tumor development will quickly bring about subtle, distinctive treatments.

Read e-book online Gene Cloning: The Mechanics of DNA Manipulation PDF

This ebook was once originallyconceived within the shape ofa moment version ofa quantity released in 1980 in Chapman and Hall's 'OutllneStudies in Biology' sequence and entitled Genetic Engineering - Cloning DNA. It very quickly turned obvious that with the impression ofrecombinant DNA options being feIt in such a lot of components ofblology, it used to be going to be difficultifnotimpossible to keepthe bookwithin the gap confines of those little monographs.

Cellular In Vitro Testing: Methods and Protocols by John Haycock, Arti Ahluwalia, J. Malcolm Wilkinson PDF

Becoming cells in second less than static stipulations has lengthy been the best of telephone tradition, regardless of this system no longer being consultant of the complicated in vivo setting. using animal versions additionally has transparent moral and clinical boundaries, and more and more the 3Rs (replacement, refinement, relief) when it comes to animal types are being built-in into the modern day clinical perform.

Extra info for Fluorescence In Situ Hybridization (FISH) - Application Guide

Example text

12. 13. 14. 15. 16. 17. T. Liehr, F. Pellestor different probes on the same slide using smaller coverslips. The amount of probe/probe-solution must be reduced according to the coverslip size. Incubate slides for 1–3 nights at 37°C in a humid chamber. 2% Tween (RT, 100 ml coplin jar). Go on with step 8 (smaller and/or more sensible probes) or 9 (larger and/or more stable probes). Post-wash the slides for 3 × 5 min in formamide solution (45°C) followed by 3 × 5 min in 2× SSC (37°C) in a 100 ml coplin jar, with gentle agitation.

4. Pellet the DNA by centrifugation at 15,000 rpm for 15 min, discard the supernatant, and dry the DNA pellet at room temperature (RT) or using a speed vac. 5. Dissolve the pellet from step 4 in 20 ml of hybridization buffer, vortex, and spin down. 6. Denature the probe solution at 75°C for 5 min, and do a pre-hybridization step at 37°C for 30 min if repetitive probe is not used. 2 Slide Pretreatment In a conventional FISH approach, pretreatment of the slides with pepsin followed by postfixation with formalin buffer is required to reduce the background.

The primers are annealed in situ to their denatured complementary DNA target sequences, and then extended by a Taq DNA polymerase in the presence of free nucleotides. The visualization of the generated fragments is enabled by the incorporation of one labeled nucleotide. The principle of PRINS is as follows (see also Fig. 2): – Fix the target DNA onto a clean microscope slide. Chromosome spreads, nuclei, extended chromatids or tissue sections can be used in PRINS reactions. – Prepare the PRINS reaction mixture incorporating the target-specific primer, the Taq DNA polymerase and its buffer, and the nucleotide mixture, including a labeled dUTP.

Download PDF sample

Fluorescence In Situ Hybridization (FISH) - Application Guide by Thomas Liehr


by William
4.0

Rated 4.70 of 5 – based on 42 votes