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文献和实验Preparation of probes for in situ hybridization
in the sequence. These free amines are chemically coupled to fluorophores after synthesis. The modified Ts should be about 10 bases apart to prevent quenching and high background. The GC content of the oligo should be around 50%, its length can be variable, but 40
Preparation of probes for in situ hybridization
after synthesis. The modified Ts should be about 10 bases apart to prevent quenching and high background. The GC content of the oligo should be around 50%, its length can be variable, but 40-50 bases (5 fluorochromes per molecule of probe) works well in most cases
by the addition of polyethylene glycol and collected by centrifugation. The DNA pellet is resuspended in a buffer containing cesium chloride and ethidium bromide, which is loaded into polyallomer tubes and subjected to ultracentrifugation overnight. The ethidium
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