by either method is usually very small, some kind of PCR amplification is normally used. The amplified products can be used for FISH (reverse painting) as described in Support Protocol 3 or can be cloned for further analysis. This allows an assessment of the purity of the chromosome fragments and is an essential step if a library is to be constructed. The DOP-PCR method described in Basic Protocol 3 was initially devised as a rapid and efficient method of amplifying microdissected chromosomes without requiring excessive technical expertise. It has worked successfully in a number of laboratories, and has been shown to give strong signals when DNA amplified from flow sorted or microdissected chromosomes is hybridized back to normal metaphase chromosomes by fluorescence in situ hybridization (FISH). For FISH analysis of PCR products, see Support Protocol 3.

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