By Graeme I. Murray, Stephanie Curran
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Extra info for Laser Capture Microdissection (Methods in Molecular Biology, v293)
Agarose gel electrophoresis equipment, 100 bp and 1 Kb DNA ladder. Fig. 1. (opposite page) Schematic overview of the principle of CGH. Differentially labeled test and normal reference DNA are co-hybridised to normal male target metaphases in the presence of C0t-1 DNA. Digital images for the hybridization pattern of test and reference DNA are acquired sequentially using fluorochrome-specific optical filters, and test (green)/reference (red) fluorescence ratio profiles along the axis of each chromosome are determined by computer-based image analysis packages.
1994) Extraction of DNA from paraffin-embedded tissue for analysis by polymerase chain reaction: new tricks from an old friend. Hum. Pathol. 25, 561–563. 7. 7 Frank, T. , Svoboda-Newman, S. , and Hsi, E. D. (1996) Comparison of methods for extracting DNA from formalin-fixed paraffin sections for nonisotopic PCR. Diagn. Mol. Pathol. 5, 220–224. 8. , Arends, J. , and de Goeij, A. (1999) Optimizing the APC gene mutation analysis in archival colorectal tumor tissue. Diagn. Mol. Pathol. 8, 11–19. 9.
5. Using the Primer Express software, the Tm should be 58–60°C. 6. The five nucleotides at the 3' end should have no more than two G and/or C bases. 1. 2. the primer and probe sequences were made as follows (in all cases, the first sequence is the forward PCR primer, the second one is the Taqman probe, and the third sequence is the reverse PCR primer): (1) ZNF217, 5'-GAG GCG AGG AAG AAG GTG C, 6FAM5'-CCC ATC TGA GAT GCT CAA AGT TGC GA3'TAMRA, 5'-CGG AAG CTG GCA GCA TTT T' and (2) ` globin, 5'-CAA GAA AGT GCT CGG TGC CT, 6FAM5'-GTC CAG GTG AGC CAG GCC ATC ACT A-3'VIC, 5'-GCA AAG GTG CCC TTG AGG T.
Laser Capture Microdissection (Methods in Molecular Biology, v293) by Graeme I. Murray, Stephanie Curran