By Diponkar Banerjee (auth.), Diponkar Banerjee, Sohrab P. Shah (eds.)
The earliest descriptions of human chromosomes initiated the genomics revolution that's now upon us. Array Comparative Genomic Hybridization: Protocols and Applicationsexplores the scope of what's now attainable so far as documenting abnormalities linked to numerous human cancers. whereas the expertise for interrogating the human genome keeps to conform, refinement of comparative genomic hybridization (CGH) utilizing array CGH and similar applied sciences have supplied huge, immense perception into human cancers at a cheap scale. Written within the profitable Methods in Molecular Biology™ sequence structure, chapters contain introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, with no trouble reproducible protocols, and notes on troubleshooting and fending off recognized pitfalls.
Authoritative and simply available, Array Comparative Genomic Hybridization: Protocols and Applicationsprovides researchers with well-honed methodologies to profit those innovations for his or her personal use in learn or medical diagnostic laboratories.
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Extra resources for Array Comparative Genomic Hybridization: Protocols and Applications
2 Ultradense Array CGH and Discovery of Micro-Copy Number… 17 Obtaining reproducible, high-resolution copy number data with high sensitivity and few false positives is the primary objective for any high throughput experimental study. According to the most comprehensive analysis published so far, reproducibility of replicate experiments of aCGH for finding CNV in DNA from normal healthy individuals is <70% (9) and may decrease even further when cancer biopsy samples are analyzed or when test DNA is subjected to whole genome amplification (WGA) prior to labeling and hybridization (8) .
Magnetic stir plate (2×) and stir bars (2×). Single-channel pipettes (P10, P20, P200, P1000). Sterile, low binding, barrier tips 100, 200, 20, and 10 μl. Ice bucket. Timer. Circulating water bath. Heat block. Nuclease-free Eppendorf tubes (autoclaved). MICROTEST Flat Bottom Polystyrene, non-tissue culture treated 96 well plates (Falcon #351172) for PicoGreen Assay. Powder-free gloves (latex) (see Note 1). Sterile tweezers. 2. General Lab Reagents Molecular biology grade water from Milli-Q Synthesis system (Millipore) (autoclaved).
T compound avoiding bubbles. 8. T has solidified). 9. T block containing the biopsy on to the cryostat (set between −25 and −30 °C) and proceed with cryosectioning of 5 μm sections (standard pathology procedure). 10. Mount 5 μm sections on glass slides and proceed with H&E staining (standard pathology protocol). 11. Have the H&E slides evaluated by a pathologist. Specimens containing at least 70% tumor cells are accepted for sorting experiments (see Note 4 and 5). 22 E. Przybytkowski et al. 2. Preparation of Biopsies for Sorting Nuclei (See Note 6) 1.
Array Comparative Genomic Hybridization: Protocols and Applications by Diponkar Banerjee (auth.), Diponkar Banerjee, Sohrab P. Shah (eds.)